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"Endometriosis merupakan penyakit pada sistem reproduksi wanita yang ditandai dengan tumbuhnya jaringan endometrium di luar rongga uterus. Endometriosis memengaruhi sistem reproduksi salah satunya dengan menyebabkan penurunan kualitas oosit akibat terjadinya apoptosis pada sel granulosa di dalam folikel. Apoptosis pada sel granulosa diketahui diaktivasi melalui jalur intrinsik yang dipengaruhi oleh protein BAX (pro-apoptosis) dan BCL-2 (anti-apoptosis). Penelitian ini bertujuan untuk mengetahui ekspresi mRNA bcl-2 dan bax pada sel granulosa penderita endometriosis melalui metode real-time PCR yang kemudian diuji secara statistik dengan menggunakan Uji-t. Hasil penelitian menunjukkan bahwa ekspresi mRNA bax/bcl-2 pada wanita penderita endometriosis menunjukkan perbedaan yang signifikan (p < 0,05) dibandingkan pada wanita tanpa endometriosis.

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Endometriosis is a disease in female reproductive system which marked by the present of endometrium tissue outside the uterus cavity. Endometriosis affects the reproductive system by decreasing oocyte quality as an impact of granulosa cells apoptosis in the follicle. Apoptosis in granulosa cells has been known activated through intrinsic pathway which is influenced by BAX (pro-apoptosis) and BCL-2 (anti-apoptosis) proteins. This research was conducted to know the mRNA expression of bcl-2 and bax in granulosa cells of endometriosis patients using real-time PCR and statistic tests (T-test). The result shows that there is significance difference (p < 0,05) of bax/bcl-2 expression between granulosa cells of endometriosis patients and granulosa cells in women without endometriosis."

"Pengantar: Endometriosis merupakan salah satu penyebab infertilitas dan menjadi indikasi fertilisasi in vitro (FIV). Laju apoptosis dan stress oksidatif yang tinggi pada pasien endometriosis diyakini menimbulkan efek negatif terhadap peluang keberhasilan FIV. Namun, pengaruh endometriosis terhadap keberhasilan FIV menunjukkan bukti yang inkonsisten dan belum banyak studi yang menilai langsung efek endometriosis terhadap kualitas oosit sebagai parameter keberhasilan FIV.Tujuan: Untuk menilai laju apoptosis pada sel granulosa pasien endometriosis dibanding pasien non-endometriosis melalui rasio ekspresi mRNA BAX/BCL-2 dan menilai korelasinya dengan kualitas oosit yang didapatkan saat petik ovum.Hasil: Sampel didapatkan dari 15 subjek dengan endometriosis dan 15 subjek kontrol. Dosis rekombinan FSH total yang diterima pada kelompok endometriosis untuk stimulasi ovarium lebih tinggi dibandingkan kelompok kontrol (p=0.005). Terdapat perbedaan bermakna kadar ekspresi BAX (p=0.029) dan BCL-2 (p<0.001) pada kedua kelompok, tetapi perbedaan rasio keduanya tidak signifikan (p=0.787). Korelasi antara rasio BAX/BCL-2 dengan parameter kualitas oosit tidak menunjukkan hubungan bermakna di kedua kelompok.Kesimpulan: Tidak terdapat perbedaan signifikan pada rasio kadar BAX/BCL-2 di kedua kelompok dan tidak ditemukan hubungan bermakna antara rasio tersebut dengan kualitas oosit. 

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Introduction: Endometriosis is one of common conditions causing infertility and an indication to undergo in vitro fertilization (IVF). High apoptosis rate and oxidative stress in patient with endometriosis is believed to cause negative effect on IVF success rate. However, there has been conflicting results on endometriosis effect to IVF success and there have been limited studies that directly assess endometriosis and its effect on oocyte quality.

Aim: To assess apoptosis rate on granulosa cells in patients with endometriosis compared to non-endometriosis patients through mRNA BAX/BCL-2 ratio and how it correlates with oocyte quality collected during ovum pick up.

Results: Samples were collected from 15 subjects with endometriosis and 15 control subjects. Total dose of recombinant FSH received by endometriosis group is significantly higher compared to control (p=0.005). There is difference in BAX level (p=0.029) and BCL-2 level (p<0.001) in both groups. However, the ratio does not differ significantly (p=0.787). No significant correlation is found in BAX/BCL-2 ratio and any of the oocyte quality parameters.

Conclusion: We found no significant difference in BAX/BCL-2 ratio between endometriosis and control group as well as significant correlation between the ratio and oocyte quality."

"Latar Belakang. Endometriosis ditandai dengan pertumbuhan jaringan endometrium di luar uterus, salah satunya disebabkan oleh disregulasi apoptosis sel yang memicu ketahanan sel ektopik. Asam galat dan turunannya pada beberapa penelitian mampu menghambat karsinogenesis pada beberapa cell line kanker. Penelitian kami terdahulu membuktikan asam galat dan turunannya dapat menekan proliferasi sel dan meningkatkan apoptosis sel endometriosis in vitro, namun efeknya terhadap mekanisme jalur apoptosis instrinsik belum di buktikan. Metode. Sel endometriosis berasal dari jaringan endometrium pasien laparaskopi, diisolasi secara enzimatis dan dikultur primer. Sel kultur diberi perlakuan asam galat, heptil galat, oktil galat dengan dosis 51,2 μg/ml, 102,4 μg/ml dan 153,6 μg/ml selama 48 jam, dilanjutkan induksi 10 ng/ml LPS selama 24 jam . Kelompok kontrol hanya di induksi LPS tanpa perlakuan. Ekspresi relatif mRNA Bax, Bcl-2, dan Caspase-3 dinilai dengan qRT-PCR. Hasil. Peningkatan tertinggi ekspresi mRNA Bax dan penekanan tertinggi ekspresi mRNA Bcl-2 pada oktil galat dosis 153,6 μg/ml. Peningkatan ekspresi mRNA Bax, dan penurunan ekspresi mRNA Bcl-2 akan di ikuti dengan peningkatan ekspresi mRNA Caspase-3. Secara statistik tidak terdapat perbedaan bermakna antara kelompok perlakuan dengan ekspresi mRNA Bax, Bcl-2, dan Caspase-3 (p > 0,05). Kesimpulan. Oktil galat, asam galat, dan heptil galat memiliki efek potensial pada mekanisme apoptosis intrinsik.

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Background. Endomeriosis characterized by the presence of extrauterine endometrial tissue, one of which caused by disregulation of apoptosis that contribute of endometrial ectopic survival. Our previous research has proven that gallic acid and its derivatives can suppress proliferation and induce apoptosis endometriosis cell in vitro. However, the effect of gallic acid and its derivatives on apoptosis intrinsic pathway mechanism is not proven yet. Method. Endometriosis cell from endometriosis patiens who had undergone laparascopy surgery were isolated by enzimatic reaction and primary cultured. Cultured cells treated by gallic acid, heptyl gallate and octyl gallate each with dosage 51.2 μg/ml, 102.4 μg/ml, 153.6μg/ml for 48 hours, than induced by LPS 10 ng/ml for 24 hours. Parameter research was assessed by qRT-PCR for mRNA expression of Bax, Bcl-2, Caspase-3. Result. Octyl gallate showed more effect to induce apoptosis intrinsic . Endometriosis cell were treated with octyl gallate shown increases of Bax and Caspase3 mRNA expression than decrease of Bcl-2 mRNA expression. Statistically, mean differences are not significant between treatment groups and mRNA expression (p > 0.05). Conclusion. This study exhibited that octyl gallate has a more potential effect on apoptosis intrinsic in endometriosis cell cultures followed by gallic acid and heptyl gallate and their potency as treatment for endometriosis."

"Endometriosis merupakan penyakit pada sistem reproduksi wanita yang ditandai dengan tumbuhnya jaringan endometrium di luar rongga uterus. Endometriosis memengaruhi sistem reproduksi salah satunya dengan menyebabkan penurunan kualitas oosit. Regulasi homon FSH dan LH pada sel granulosa penderita endometriosis tidak berada pada kondisi normal, yaitu kadar estrogen yang tinggi sebelum ovulasi dan adanya indikasi kekurangan reseptor LH sehingga menyebabkan penundaan pelonjakan LH yang akan memengaruhi proses pematangan oosit. Hal ini dipengaruhi oleh regulasi hormon FSH dan LH yang memiliki korelasi positif terhadap reseptornya, yaitu fshr dan lhr. Penelitian ini bertujuan untuk mengetahui ekspresi gen fshr dan lhr pada sel granulosa penderita endometriosis melalui metode real-time PCR yang kemudian diuji secara statistik dengan menggunakan Uji-t. Hasil penelitian menunjukkan bahwa ekspresi mRNA fshr/lhr pada wanita penderita endometriosis lebih tinggi dibandingkan dengan kondisi normal, walaupun perbedaannya tidak signifikan. Perbedaan rata-rata ekspresi mRNA fshr/lhr antara penderita endometriosis dan wanita normal adalah 0,084.

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Endometriosis is a disease of the female reproductive system marked with the growth of endometrial tissue outside the uterine cavity. Endometriosis affects the reproductive system by decreasing oocyte quality as an impact of hormone alteration. Regulatory hormones FSH and LH in the granulosa cells of endometriosis patients are not on normal conditions, estrogen level is very high before ovulation and there is indication of deficiency LH receptor in endometriosis that causes delayed of LH surge. This is influenced by FSH and LH hormone regulation that has a positive correlation to its receptor, fshr and lhr. This research was conducted to know the mRNA expression of fshr and lhr in granulosa cells of endometriosis patients using real time PCR and statistic test T test. The result shows that expression of fshr lhr ini endometriosis patients are higher than in women without endometriosis, although not significance. Mean difference of fshr lhr mRNA expression between endometriosis patients and normal conditions is 0,084."

"Latar Belakang: Salah satu hipotesis yang menjelaskan hubungan endometriosis dengan infertilitas adalah endometriosis diyakini menyebabkan gangguan fisiologi ovarium, salah satunya dengan mempengaruhi folikulogenesis yang menyebabkan penurunan kualitas oosit. Oosit memainkan peran penting dalam mengatur dan mendukung pertumbuhan folikel, melalui produksi faktor pertumbuhan oosit. Beberapa faktor pertumbuhan telah diidentifikasi pada oosit manusia, termasuk growth differentiation factor-9 GDF-9 . Namun, sampai saat ini penelitian mengenai ekspresi GDF-9 pada sel granulosa pada wanita infertil dengan endometriosis masih belum banyak dilakukan. Tujuan: Untuk mengetahui ekspresi mRNA GDF-9 pada sel granulosa pasien endometriosis yang menjalani FIV dan untuk mencari adanya korelasi antara ekspresi GDF-9 dengan kualitas oosit. Metode: Penelitian potong lintang ini dilakukan di Klinik IVF Yasmin RSCM dan Klinik Sander B di Jakarta pada bulan Juli 2014 - Juli 2017. Sebanyak 50 sampel terdiri atas 25 wanita dengan endometriosis dan 25 kontrol. Sampel sel granulosa dikumpulkan pada saat petik oosit. Ekspresi mRNA GDF-9 dinilai menggunakan real time PCR. Hasil: Terdapat penurunan jumlah ambilan oosit, jumlah oosit matur dan skor morfologi oosit pada kelompok pasien dengan endometriosis dan bermakna secara statistik. Ekspresi GDF-9 secara kuantitatif lebih rendah pada kelompok endometriosis dibandingkan dengan kontrol 5.05 0.00002 ndash; 3523 ng/ l vs 81.93 1,47 ndash; 32450 ng/ l; p=0,01 . Pada penelitian ini tidak didapatkan korelasi antara ekspresi GDF-9 dan kualitas oosit dari skor morfologi oosit dan laju fertilisasi. Kesimpulan: Ekspresi GDF9 lebih rendah pada kelompok endometriosis dibandingkan kelompok kontrol. Namun, kami tidak menemukan korelasi antara ekspresi GDF-9 dengan kualitas oosit. Dibutuhkan studi dengan besar sampel yang lebih besar untuk mengkonfirmasi apakah perubahan ekspresi GDF-9 memiliki korelasi dengan kualitas oosit serta untuk membuktikan apakah GDF-9 dapat digunakan sebagai penanda molekuler baru untuk memprediksi kompetensi perkembangan oosit.

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Background: One of the hypothesis that explains the association between endometriosis and infertility is that endometriosis is believed to cause ovarian physiology disturbances, one of them by affecting folliculogenesis that cause decreased oocyte quality. The oocyte plays an important role in regulating and promoting follicle growth, by the production of oocyte growth factors. Several growth factors have been identified in human oocytes, including growth differentiation factor-9 GDF-9. However the studies on GDF-9 expression in granulosa cells of infertile women with endometriosis are sparse.

Objective: To investigate the expression of GDF-9 mRNA in granulosa cells of endometriosis patients undergoing IVF and to find the correlation between GDF-9 expression and oocyte quality.

Method: This cross sectional study was done at Yasmin IVF Clinic and dr. Sander B Clinic Jakarta in July 2014 - July 2017. A total fifty samples of 25 womens with endometriosis and 25 controls were included. We collect the granulosa cells sample at the time of oocyte retrieval. GDF-9 mRNA expression were investigated by Real-Time PCR.

Result: The number of oocytes retrieved, mature oocytes and the oocyte morphology score were lower in the group of patients with endometriosis and this was statistically significant. GDF-9 mRNA expression levels was quantitatively lower in endometriosis groups compared to control 5.05 0.00002 ndash; 3523 ng/ l vs 81.93 1,47 ndash; 32450 ng/ l; p=0,01. However, we did not find any correlation between GDF-9 expression levels and oocyte quality from oocyte morphology score and fertilization rate.

Conclusion: GDF9 mRNA level was lower in endometriosis group compared to control group. However, we did not find correlation between individual GDF-9 level and oocyte quality. Large-sample studies were needed to confirm whether the expression of GDF-9 had a correlation with oocyte quality as well as to prove whether GDF-9 could be used as a new molecular marker to predict the oocyte developmental competence."

"Pendahuluan: Endometriosis merupakan suatu kelainan jinak ginekologi yang dapat mengalami transformasi menjadi kanker. Stres oksidatif diduga berperan dalam perkembangan penyakit endometriosis. Gen supresor tumor ARID1A banyak ditemukan termutasi dan inaktif pada kanker ovarium yang berhubungan dengan endometriosis. Tujuan penelitian adalah untuk menganalisis peran stres oksidatif terhadap ekspresi gen supresor tumor ARID1A dalam transformasi endometriosis menjadi ganas. Metoda: Penelitian dimulai dengan 10 sampel jaringan kanker ovarium, 10 sampel endometriosis dan3 jaringan endometrium eutopik sebagai kontrol yang diisolasi mRNA dan proteinnya. Analisis ekspresi gen ARID1A pada tingkat mRNA dilakukan dengan pemeriksaan RT-qPCR dan pada tingkat protein dengan ELISA. Pada sel endometriosis dan kanker ovarium dilakukan analisis stres oksidatif dengan pemeriksaan aktivitas antioksidan MnSOD dan pemeriksaan kadar MDA sebagai salah bukti kerusakan salah satu komponen sel. Setelah itu dilakukan uji eksperimental pada kultur sel endometriosis dan endometrium eutopik sebagai kontrol. Kedua sel kultur diinduksi dengan H2O2 konsentrasi 0 nM, 100 nM, dan 1000 nM. Analisis dilakukan terhadap ketahanan hidup sel, kadar ROS dan ekspresi gen ARID1A pada tingkat mRNA dan protein. Hasil: Efek induksi H2O2 dalam menekan ekspresi gen ARID1A sel endometriosis dan sel endometrium eutopik pada tingkat mRNA dan protein, bermakna, meskipun pada kanker ovarium tidak bermakna pada penelitian ini. Kesimpulan: Stres oksidatif berperan dalam menekan ekspresi gen supresor tumor ARID1A ditingkat mRNA dan protein pada endometriosis.

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Introduction: Endometriosis as a gynecologic benign lesion, can transform itself into cancer. Oxidative stress is considered as an important factor in endometriosis development. Studies found that ARID1A as tumor suppressor gene, was frequently mutated and inactivated in endometriosis associated ovarian cancer. The aim of the study is to analyze the role of oxidative stress on ARID1A expresion in endometriosis malignant transformation.

Methods: This study started with ten samples of ovarian cancer, ten samples of endometriosis, and 3 samples of eutopic endometrioid tissues as control. They were analyzed for the expression of ARID1A by RT-qPCR and ELISA, then analyzed for the activity of MnSOD as antioxidant enzyme and level of malondialdehyde as one of the oxidative stress damage effect evidence on cell's components. The second part of the study was experimental study on cultured eutopic endometrial and endometriosis cells. They were induced by H2O2 of 0, 100, and 1000 nM concentration. Analysis of the expression of ARID1A by RTqPCR and ELISA, and the DCFH-DA for the level of Reactive oxygen species were done.

Result: The impact of the H2O2 induction in repressing ARID1A gene expression on the endometriosis as well on the eutopic endometrium cells are significant, but not on the ovarian cancer in this study.

Conclusion: Oxidative stress has a role in repressing the expression of ARID1A gene at the mRNA and protein levels on the endometriosis."

"[ABSTRAKLatar Belakang: Kanker prostat adalah kanker yang paling umum pada pria. Kanker terjadi karena hilangnya kontrol atas proliferasi sel dan apoptosis sehingga sel berproliferasi terus menerus tanpa ada kematian sel. Apoptosis diregulasi oleh beberapa protein tertentu diantaranya protein keluarga Bcl-2 dan protein kanal. Perkembangan kanker prostat memerlukan transformasi dari sel epitel yang normal menjadi sel ganas yang kehilangan kemampuan untuk mengakumulasi zinc. Salah satu efek utama zinc adalah mencegah pertumbuhan sel kanker prostat dengan menginduksi apoptosis dengan memfasilitasi proses pembentukan pori Bax yang memulai apoptogenesis mitokondria. Selain keluarga Bcl-2, VDAC1 juga berperan penting dalam proses apoptosis. Beberapa penelitian menyatakan Bcl-2 mempunyai kaitan erat dengan VDAC1 terkait proses apoptosis dan protein pro-apoptotik Bax juga secara langsung berinteraksi dengan VDAC yang kemudian menginduksi keluarnya sitokrom c dari membran mitokondria.Tujuan: Mengevaluasi ekspresi mRNA dari gen mengkode keluarga protein Bcl-2 (Bax dan Bcl-2) dalam proses apoptogenesis pada galur sel kanker prostat yg diinduksi oleh zinc; Mengevaluasi ekspresi mRNA dari gen VDAC1 dalam proses apoptogenesis pada galur sel kanker prostat yang diinduksi oleh zinc; Menganalisis hubungan antara ekspresi VDAC1 dengan protein keluarga Bcl-2 pada apoptogenesis galur sel kanker prostat.Desain: Penelitian ini menggunakan eksperimental in vitro dan analisis statistikMetode: Untuk memperbanyak galur sel kanker prostat (PC3) dilakukan kultur sel, kemudian diberi perlakuan dengan tiga kelompok (kontrol, zinc 20 μM dan staurosporin 0,16 μM). Selanjutnya dilakukan isolasi RNA dan elektroforesis RNA untuk mengetahui keutuhan RNA. Terakhir dilakukan qRT PCR yang kemudian datanya dianalisis secara statistika.Hasil: Ekspresi Bax, Bcl-2 dan VDAC1 pada galur sel kanker prostat (PC-3) yang diberi perlakuan zinc mengalami penurunan dibandingkan dengan kontrol (tidak diberi perlakuan). Akan tetapi penurunan ekspresi tersebut tidak bernilai signifikan karena nilai p > 0,05 (nilai signifikansi Bax = 0,309; nilai signifikansi Bcl-2 = 0,236; nilai signifikansi VDAC1 = 0,437). VDAC1 mempunyai korelasi yang signifikan (p < 0,05) dengan Bax (p = 0,01) dibandingkan dengan Bcl-2 (p = 0,118).Kesimpulan: Terjadi perubahan ekspresi pada setiap gen (Bax, Bcl-2 dan VDAC1) pada galur sel kanker prostat yang diberi perlakuan zinc dengan yang tidak diberi perlakuan, akan tetapi tidak bernilai signifikan. VDAC1 mempunyai korelasi yang bermakna dengan Bax dan mempunyai korelasi yang tidak bermakna dengan Bcl-2.

ABSTRACT

Background: Prostate cancer is the most common cancer in men. Cancer occurs due to loss control of cell proliferation and apoptosis thus continuously proliferating cells without cell death. Apoptosis is regulated by specific proteins including Bcl-2 family proteins and channel proteins. The development of prostate cancer requires the transformation of normal epithelial cells into malignant cells that lose the ability to accumulate zinc. One of the main effects of zinc is to prevent the growth of prostate cancer cells by inducing apoptosis by facilitating the process of pore formation Bax that started apoptogenesis mitochondrial. In addition to Bcl-2 family, VDAC1 also plays an important role in the process of apoptosis. Some studies suggest Bcl-2 has close links with related VDAC1 apoptosis and pro-apoptotic protein Bax also directly interact with VDAC which then induces the release of cytochrome c from the mitochondrial membrane.

Objective: To evaluate the expression of mRNA of the gene encoding the Bcl-2 family proteins (Bax and Bcl-2) in the process apoptogenesis on prostate cancer cell line that is induced by zinc; Evaluate the mRNA expression of genes in the process VDAC1 apoptogenesis on prostate cancer cell line induced by zinc; Analyzing the relationship between the expression of VDAC1 with Bcl-2 family proteins in prostate cancer cell lines apoptogenesis.

Design: This study used an experimental in vitro and statistical analysis

Methods: To reproduce the prostate cancer cell lines (PC3) performed cell culture, then treated with three groups (control, zinc 20 μM and staurosporin 0,16 μM). Furthermore, the isolation of RNA and RNA electrophoresis to determine the integrity of the RNA. Recently performed qRT PCR and the data were analyzed statistically.

Results: The expression of Bax, Bcl-2 and VDAC1 on prostate cancer cell line (PC-3) were treated with zinc decreased than the control (untreated). However, a decrease in the expression of no significant value because the value of p > 0.05 (Bax significant value = 0.309; the value of the significance of Bcl-2 = 0.236; VDAC1 significant value = 0.437). VDAC1 has a significant correlation (p < 0.05) with Bax (p = 0.01) than Bcl-2 (p = 0.118).

Conclusion: There is a change in the expression of each gene (Bax, Bcl-2 and VDAC1) in prostate cancer cell lines that treated with zinc than untreated, but no significant value. VDAC1 has a significant correlation with Bax and had no significant correlation with Bcl-2.;Background: Prostate cancer is the most common cancer in men. Cancer occurs due to loss control of cell proliferation and apoptosis thus continuously proliferating cells without cell death. Apoptosis is regulated by specific proteins including Bcl-2 family proteins and channel proteins. The development of prostate cancer requires the transformation of normal epithelial cells into malignant cells that lose the ability to accumulate zinc. One of the main effects of zinc is to prevent the growth of prostate cancer cells by inducing apoptosis by facilitating the process of pore formation Bax that started apoptogenesis mitochondrial. In addition to Bcl-2 family, VDAC1 also plays an important role in the process of apoptosis. Some studies suggest Bcl-2 has close links with related VDAC1 apoptosis and pro-apoptotic protein Bax also directly interact with VDAC which then induces the release of cytochrome c from the mitochondrial membrane.

Objective: To evaluate the expression of mRNA of the gene encoding the Bcl-2 family proteins (Bax and Bcl-2) in the process apoptogenesis on prostate cancer cell line that is induced by zinc; Evaluate the mRNA expression of genes in the process VDAC1 apoptogenesis on prostate cancer cell line induced by zinc; Analyzing the relationship between the expression of VDAC1 with Bcl-2 family proteins in prostate cancer cell lines apoptogenesis.

Design: This study used an experimental in vitro and statistical analysis

Methods: To reproduce the prostate cancer cell lines (PC3) performed cell culture, then treated with three groups (control, zinc 20 μM and staurosporin 0,16 μM). Furthermore, the isolation of RNA and RNA electrophoresis to determine the integrity of the RNA. Recently performed qRT PCR and the data were analyzed statistically.

Results: The expression of Bax, Bcl-2 and VDAC1 on prostate cancer cell line (PC-3) were treated with zinc decreased than the control (untreated). However, a decrease in the expression of no significant value because the value of p > 0.05 (Bax significant value = 0.309; the value of the significance of Bcl-2 = 0.236; VDAC1 significant value = 0.437). VDAC1 has a significant correlation (p < 0.05) with Bax (p = 0.01) than Bcl-2 (p = 0.118).

Conclusion: There is a change in the expression of each gene (Bax, Bcl-2 and VDAC1) in prostate cancer cell lines that treated with zinc than untreated, but no significant value. VDAC1 has a significant correlation with Bax and had no significant correlation with Bcl-2., Background: Prostate cancer is the most common cancer in men. Cancer occurs due to loss control of cell proliferation and apoptosis thus continuously proliferating cells without cell death. Apoptosis is regulated by specific proteins including Bcl-2 family proteins and channel proteins. The development of prostate cancer requires the transformation of normal epithelial cells into malignant cells that lose the ability to accumulate zinc. One of the main effects of zinc is to prevent the growth of prostate cancer cells by inducing apoptosis by facilitating the process of pore formation Bax that started apoptogenesis mitochondrial. In addition to Bcl-2 family, VDAC1 also plays an important role in the process of apoptosis. Some studies suggest Bcl-2 has close links with related VDAC1 apoptosis and pro-apoptotic protein Bax also directly interact with VDAC which then induces the release of cytochrome c from the mitochondrial membrane.

Objective: To evaluate the expression of mRNA of the gene encoding the Bcl-2 family proteins (Bax and Bcl-2) in the process apoptogenesis on prostate cancer cell line that is induced by zinc; Evaluate the mRNA expression of genes in the process VDAC1 apoptogenesis on prostate cancer cell line induced by zinc; Analyzing the relationship between the expression of VDAC1 with Bcl-2 family proteins in prostate cancer cell lines apoptogenesis.

Design: This study used an experimental in vitro and statistical analysis

Methods: To reproduce the prostate cancer cell lines (PC3) performed cell culture, then treated with three groups (control, zinc 20 μM and staurosporin 0,16 μM). Furthermore, the isolation of RNA and RNA electrophoresis to determine the integrity of the RNA. Recently performed qRT PCR and the data were analyzed statistically.

Results: The expression of Bax, Bcl-2 and VDAC1 on prostate cancer cell line (PC-3) were treated with zinc decreased than the control (untreated). However, a decrease in the expression of no significant value because the value of p > 0.05 (Bax significant value = 0.309; the value of the significance of Bcl-2 = 0.236; VDAC1 significant value = 0.437). VDAC1 has a significant correlation (p < 0.05) with Bax (p = 0.01) than Bcl-2 (p = 0.118).

Conclusion: There is a change in the expression of each gene (Bax, Bcl-2 and VDAC1) in prostate cancer cell lines that treated with zinc than untreated, but no significant value. VDAC1 has a significant correlation with Bax and had no significant correlation with Bcl-2.]"

"B-cell leukaemia/lymphoma-2 Bcl-2 dan Bcl-2-associated X protein Bax merupakan anggota dari Bcl-2 family yang berperan dalam meregulasi apoptosis. Apoptosis penting dalam perkembangan manusia. Terganggunya kejadian apoptosisakan memberikan efek terhadap keadaan fisiologis manusia, diantaranya gangguan reproduksi. Salah satu gangguan reproduksi yang dialami oleh wanita usia reproduktif yaitu Sindrom Ovarium Polikistik (SOPK). Wanita penderita SOPK memiliki jumlah folikel yang lebih banyak dibandingkan wanita tanpa SOPK. Penelitian bertujuan untuk mengetahui ekspresi mRNA bax dan bcl-2 pada wanita SOPK serta mengetahui korelasi rasio ekspresi mRNA bax terhadap bcl-2 dengan rasio fertilisasi. Sel granulosa untuk penelitian didapatkan dari 18 wanita penderita SOPK dan 10 wanita tanpa SOPK yang sedang menjalani program FIV. Hasil penelitian menunjukkan tidak ada perbedaan yang berarti antara rasio ekspresi mRNA bax/bcl-2 wanita penderita SOPK dan tanpa SOPK p > 0,05. Korelasi yangkuat R = 0,525 ditemukan antara rasio ekspresi bax/bcl-2 dengan rasio fertilisasi.

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B cell leukemia lymphoma 2 Bcl 2 and Bcl 2 associated X protein Bax are members of the Bcl 2 family that play a role regulating apoptosis. Apoptosis plays an important role in human development. Disruption of apoptosis will have an effect on the physiological state of humans, including reproductive disorders. One of there productive disorders experienced by women in reproductive age is Polycystic Ovary Syndrome (PCOS). Women with PCOS have a higher number of follicles than women without PCOS.

The aim of this study was to find out the expression of baxand bcl 2 mRNA expression in PCOS women and to know the correlation of bax mRNA expression ratio to bcl 2 with fertilization ratio. Granulosa cells for the study were obtained from 18 women with PCOS and 10 women without PCOS undergoing IVF program. The results showed no significant difference between the expression ratio of bax bcl 2 mRNA women with PCOS and without PCOS p 0.05. A strong correlation R 0.525 was found between the expression ratio of bax bcl 2and the fertilization rate."

"Endometriosis adalah kelainan ginekologis yang dimanifestasikan dengan adanya kelenjar dan sel endometrium yang berkembang di luar uterus. Endometriosis merupakan penyakit multifaktorial dimana faktor genetik dan lingkungan berinteraksi menyebabkan timbulnya penyakit ini. Beberapa penelitian telah melaporkan bahwa endometriosis merupakan penyakit yang terkait dengan hormon estrogen. Mekanisme kerja estrogen ditentukan oleh kuantitas dan aktivitas reseptor estrogen. Namun demikian analisa variasi genetik, ekspresi dan aktivitas estrogen reseptor sampai saat ini belum banyak diketahui. Tujuan Penelitian ini adalah untuk menganalisa variasi genetik, ekspresi mRNA dan aktivasi ER pada jaringan endometriosis. Alel gen reseptor estrogen REα dan REβ dari 83 sampel penderita endometriosis dibandingkan dengan 76 kontrol menggunakan metoda PCR RFLP. Pengukuran ekspresi mRNA dari 18 jaringan penderita endometriosis dan 18 kontrol dilakukan dengan menggunakan metoda kuantitatif Real Time PCR (qRT-PCR). Pengukuran kadar estrogen serum (E2) dilakukan dengan metoda ELISA. Deteksi aktivasi ER dilakukan dengan uji fosforilasi reseptor estrogen β (serin 105) dengan metoda Western Blot. Hasil uji Chi-square ditemukan bahwa frekuensi alel A (normal) dan alel G (mutan) pada gen REα SNP rs9340799 dalam populasi berbeda bermakna (p=0,012) dan OR 1,772 dan kedua frekuensi alel dari hasil uji keseimbangan menurut Hardy-Weinberg berbeda bermakna (p = 0,003). Frekuensi alel (normal dan mutan) dalam populasi REα SNP rs2234693 tidak menunjukkan perbedaan bermakna dan seimbang dalam populasi. Frekuensi genotip pada SNP REβ rs4986938 pada endometriosis dibandingkan kontrol berbeda bermakna (p=0,015) dan OR 0,311 dengan populasi seimbang. Menurut keseimbangan Hardy-Weinberg dan frekuensi alel normal G dan alel mutan A juga berbeda bermakna (p=0,034) dan OR =0,438. Hasil pengukuran ekspresi mRNA menunjukkan terjadi peningkatan ekspresi ERβ 49,52 kali dibanding kontrol sedangkan REα tidak menunjukkan perbedaan dibandingkan kontrol. Kadar estradiol serum (E2) fase proliferasi tidak menunjukkan perbedaan bermakna dibanding kontrol. Hasil uji Spearman menunjukkan tidak ada korelasi kadar estradiol dengan ekspresi REα dan REβ (p>0,05). Fosforilasi ERβ pada Serin 105 menunjukkan penurunan pada kelompok endometriosis dibandingkan jaringan normal dengan perbandingan nilai intensitas pita yakni 0,1 pada endometriosis dan 4,2 pada kontrol. Sebagai kesimpulan, frekuensi alel A dan G gen REα berbeda bermakna pada SNP rs9340799 dan frekuensi alel di dalam populasi tidak seimbang. Distribusi genotip normal GG dan mutan AA serta frekuensi alel G dan alel A gen REβ berbeda bermakna pada SNP rs4986938. Ekspresi (mRNA) REβ lebih tinggi secara signifikans pada kelompok endometriosis dibandingkan kontrol. Ekspresi protein Fosforilasi ERβ pada Serin 105 menunjukkan penurunan pada endometriosis dibandingkan jaringan normal.

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Endometriosis is a gynecological disorder that is manifested by the presence of endometrium glands and cells that grow and develop outside the uterus. Endometriosis is a multifactorial with genetic and environmental factors interacting to cause this disease. Several studies have reported that endometriosis is a disease associated with the hormone estrogen. The mechanism of action of estrogen depends on the quantity and activity of estrogen receptors. However, genetic variation, expression and estrogen receptor activation in endometriois patients have not been fully characterized. The aim of this study was to analyze genetic variation, ER expression and determine ER activation in endometriosis patients. This study determined the alleles of the estrogen receptor gene REα and REβ from 83 blood samples from endometriosis patients compared with 76 controls using the RFLP PCR method. Quatitative Real time PCR was used to analyze mRNA expression of REβ genes from 18 tissues with endometriosis and 18 controls. Measurement of serum estrogen (E2) levels was carried out using the ELISA method. Furthermore, the phosphorylation test of estrogen receptor (serin 105) was carried out using the Western Immunobloting method. The results of the Chi-square test found that the frequencies of the A (normal) allele and G (mutant) allele in the REα SNP gene rs9340799 in the population were significantly different (p=0.012) and OR 1.772 and the two allele frequencies from the results of the balance test according to Hardy-Weinberg were significantly different. (p = 0.003). Allele frequencies (normal and mutant) in the REα SNP population of rs2234693 did not show significant and balanced differences in the population. The genotype frequency of SNP REβ rs4986938 in endometriosis compared to control was significantly different (p=0.015) and OR 0.311 with a balanced population. According to the Hardy-Weinberg balance, the frequencies of the normal G allele and the mutant A allele were also significantly different (p=0.034) and OR=0.438. Erβ expression showed 49,52 folds increase compared to control (p=0.00), whereas ERα did not show a significat different compared to control. There was no different in serum estradiol (E2) levels compared to controls. The results of the Spearman test showed that there was no correlation between serum estradiol levels and the expression of REβ and REβ (p>0.05). Phosphorylation Ser105 of ERβ showed a decrease in the endometriosis group compared to control with a comparison of values of 0.1 and 4 2. As a conclusion, The A and G allele frequencies of the RE gen gene were significantly different in SNP rs9340799 and the allele frequencies in the population were not balanced. The distribution of normal genotypes of GG and AA mutants and the frequency of G allele and A allele of REβ gene were significantly different at SNP rs4986938. REβ (mRNA) expression was significantly higher in the endometriosis group than the control group. Phosphorylated ERβ protein expression in Serin 105 showed a decrease in endometriosis compared to normal tissue."

"Endometriosis merupakan jaringan yang ditemukan di luar uterus dan dapat menjadi faktor penyebab terjadinya infertilitas. Endometriosis dapat terjadi pada wanita umur reproduktif dengan probabilitas sebesar 10—15%. Selain itu, endometriosis dapat terjadi pada wanita yang memiliki nyeri kronis pada pelvis dengan besar probabilitas sebesar 70%. Pertumbuhan dan perkembangan jaringan endometriosis dipengaruhi oleh autofagi. Mekanisme autofagi yang terjadi pada tubuh manusia dapat dipicu dengan keberadaan HMGB1. Selain dengan HMGB1, autofagi dapat dipicu dengan adanya ekspresi gen LC3 dan BECN1. Adanya HMGB1 berpengaruh terhadap jumlah gen LC3 dan BECN1. Penelitian dilakukan dengan menganalisis sampel jaringan endometriosis pada fase proliferasi dan sekretori menggunakan metode RT-qPCR absolut dengan kurva standard. Kurva standard dibuat dengan fragmen gen HMGB1 didapatkan nilai efisiensi sebesar 88,38% dan nilai R2 sebesar 0,99618. Kedua nilai yang telah disebutkan masuk ke dalam kisaran yang dapat diterima yaitu dalam rentang 80—110% dan diatas 0,99. Sementara itu, Uji T menunjukkan perbedaan dengan hasil yang tidak signifikan (p<0,05) namun menunjukkan kecenderungan pada rata-rata kelompok. Penelitian ini juga melakukan uji Spearman’s Rank pada HMGB1 dan BECN1 dan menunjukkan korelasi positif baik itu pada kelompok sekretori maupun proliferasi dengan hasil masing-masing 0,721 dan 0,729. Sehingga dapat disimpulkan dari penelitian yang dilakukan bahwa jumlah salinan gen HMGB1 berperan dalam mekanisme autofagi dan memengaruhi tingkat autofagi pada jaringan endometriosis.

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Endometriosis is human tissue found outside the uterus and can be a contributing factor to infertility. Endometriosis can occur in women of reproductive age with a percent probability of 10—15%. In addition, it can occur in women who have chronic pelvic pain with a 70%. The growth of endometriotic tissue can be influenced by autophagy. The mechanism of autophagy that occurs in the human body can be triggered by the presence of the HMGB1 protein. In addition to the HMGB1 protein, autophagy also regulated by the presence of LC3 and BECN1 genes. The presence of HMGB1 protein affects LC3 and BECN1. The study was conducted by analyzing tissue samples of endometriosis in the proliferative and secretory phase using the RT-qPCR absolute method with standard curve. The standard curve was made by HMGB1 gene fragment and obtained an efficiency value of 88.38% and R2 value of 0.99618. The two values ​​mentioned above fall into the acceptable range, namely in the range of 80-110% and above 0.99. Meanwhile, the results the T test showed there is no significant difference (p<0,05) but showed a tendency if it compared by the group means. Spearman's Rank test showed a positive correlation between HMGB1 and BECN1 expression in both the secretory and proliferative phase with results of 0.721 and 0.729, respectively. So, it can be concluded from the research that was conducted, it was found that the expression of HMGB1 gene mRNA in the secretory phase was higher in endometrial tissue while in the proliferative phase it was higher in endometrial tissue. In addition, it can be concluded that there is a tendency for the HMGB1 gene take parts to autophagy mechanism in endometriotic tissue.

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