應用大數據分析和細胞試驗探討孕三烯酮對子宮頸癌的腫瘤抑制機轉

Exploring the tumor suppression mechnism of gestrinone in cervical cancer by big data analysis and cellular experiments

邱淮浩(Huai-Hao Ciou)

指導教授 ： 曾淑玲

中山醫學大學/醫學院/醫學研究所/博士(2022年)

https://doi.org/10.6834/csmu202200027

摘要

子宮內膜異位症是最常見的婦科疾病，被證實會增加婦科癌症的罹患風險。gestrinone臨床上被應用在子宮內膜異位症的治療，然而，gestrinone對於婦科癌細胞的作用機轉則尚未釐清，因此我們想了解經gestrinone治療對子宮內膜異位症患者婦科癌症發生率的影響，並探討gestrinone在癌細胞中的抗癌機轉。
我們首先以2004年至2013年台灣健保資料庫（LHID2000）的回顧性研究與中山醫學大學附設醫院婦產科用藥紀錄檔與癌症登錄檔分析子宮內膜異位症婦女的罹癌風險，以及gestrinone對婦科癌症發生率的影響。隨後利用選擇子宮頸癌細胞HeLa、卵巢癌細胞ES-2、子宮內膜癌細胞RL95-2、HEC-1-A及正常細胞293T，藉由MTT assay、流式細胞儀、西方墨點法與即時定量PCR觀察gestrinone對婦科癌細胞產生的相關作用與抑癌機制探討。
健保資料庫結果顯示，經1:1配對的8,330名子宮內膜異位症患者與控制組相比，其卵巢癌罹癌風險為3.88倍 (95% CI, 2.04-7.38﹔P < 0.001)，子宮內膜癌罹癌風險為3.53倍(95% CI, 1.58-7.89﹔p<0.001)﹔子宮頸癌則無顯著差異 (p = 0.249)。而健保資料庫中193名子宮內膜異位症患者與中山醫學大學附設醫院中1,540名子宮內膜異位症患者使用gestrinone後皆證明能明顯降低罹患子宮頸癌的風險 (p < 0.001)。透過後續基礎試驗則證實gestrinone的抗癌能力，能夠有效且專一性抑制子宮頸癌細胞HeLa生長，透過磷酸化JNK和C-Jun訊息傳遞路徑，進而造成P21蛋白的表現下降，最終造成細胞凋亡。
透過全國性及區域性的資料庫分析結合基礎細胞試驗都證實了子宮內膜異位用藥gestrinone，可能透過活化JNK/C-Jun減少P21表現，造成細胞凋亡來降低罹患子宮頸癌的風險，或許未來能夠遵循老藥新用的模式，做為一個子宮頸癌的治療策略。

關鍵字

子宮內膜異位症 ； 子宮頸癌 ； 孕三烯酮 ； 大數據

並列摘要

Endometriosis has been shown to increase the risk of gynecological cancers. However, the effect of gestrinone, a clinical endometriosis drug, on gynecological cancers remains unclear. This study aimed to understand the effect of gestrinone on gynecological cancers. A retrospective study was conducted using the Longitudinal Health Insurance Database 2000 of the Taiwan National Health Insurance Research Database (NHIRD) to observe the risk of gynecological cancers. Medication records from the Department of Obstetrics and Gynecology, Chung Shan Medical University Hospital CSMUH and cancer records from the Taiwan Cancer Registry were collected to analyze the correlation between gestrinone use and gynecological cancers. Subsequently, human cell lines were used to investigate the effect of gestrinone on gynecological cancers. A total of 8,330 endometriosis patients were enrolled, and analyses revealed that endometriosis patients had a higher risk of developing ovarian and endometrial cancer. However, the rate of cervical cancer was not statistically different (P = 0.249). Analyses of both the NHIRD and CSMUH databases revealed that gestrinone may reduce the risk of gynecological cancer. Cellular experiments verified the anti-cancer effects of gestrinone, which effectively and specifically inhibited the growth of HeLa cervical cancer cells, decreased P21 expression via JNK/C-Jun phosphorylation, and induced apoptosis. Combining the results of clinical database analysis and cell experiments, our findings prove that gestrinone has the potential to protect against cancer through regulation of the JNK-P21 axis. Repurposing the anti-cancer efficacy of gestrinone may be a strategy for targeted therapy in the future.

並列關鍵字

endometriosis ； cervical cancer ； gestrinone ； big data

參考文獻

Ali, A. T. (2013). "Risk factors for endometrial cancer." Ceska Gynekol 78(5): 448-459.

Androutsopoulos, V. P. and D. A. Spandidos (2018). "Anticancer pyridines induce G2/M arrest and apoptosis via p53 and JNK upregulation in liver and breast cancer cells." Oncol Rep 39(2): 519-524.

Barakat, R. R., B. N. Bundy, N. M. Spirtos, J. Bell, R. S. Mannel and S. Gynecologic Oncology Group (2006). "Randomized double-blind trial of estrogen replacement therapy versus placebo in stage I or II endometrial cancer: a Gynecologic Oncology Group Study." J Clin Oncol 24(4): 587-592.

Bermudez, A., N. Bhatla and E. Leung (2015). "Cancer of the cervix uteri." Int J Gynaecol Obstet 131 Suppl 2: S88-95.

Bogoyevitch, M. A. and B. Kobe (2006). "Uses for JNK: the many and varied substrates of the c-Jun N-terminal kinases." Microbiol Mol Biol Rev 70(4): 1061-1095.