天津医药 ›› 2020, Vol. 48 ›› Issue (8): 695-699.doi: 10.11958/20193497

• 细胞与分子生物学 • 上一篇    下一篇

下调TMEM97对卵巢癌细胞SKOV3增殖、凋亡的影响及其相关机制探讨

刘晋芳1,徐小燕2,周儒奎1,徐慧超3,高艳3,苗宇船1△   

  1. 1山西中医药大学基础医学院基础医学系(邮编030024);2中国医科大学基础医学院病理生理学教研室;3山西中医药大学研究生院
  • 收稿日期:2019-11-18 修回日期:2020-05-17 出版日期:2020-08-15 发布日期:2020-08-12
  • 通讯作者: △通信作者 E-mail:mych65@163.com E-mail:mych65@163.com
  • 作者简介:刘晋芳(1991),女,硕士,助教,主要从事肿瘤增殖与癌变方面研究
  • 基金资助:
    国家自然科学基金资助项目(81470190);山西中医药大学科技创新能力培育计划项目(2019PY-026)

Effects and mechanism of down-regulated TMEM97 on proliferation and apoptosis of #br# ovarian cancer SKOV3 cells#br#

LIU Jin-fang1, XU Xiao-yan2, ZHOU Ru-kui1, XU Hui-chao3, GAO Yan3, MIAO Yu-chuan1△   

  1. 1 Department of Basic Medical Sciences, Basic Medical College, Shanxi University of Traditional Chinese Medicine, Taiyuan 030024, China; 2 Department of Pathophysiology, Basic Medical College, China Medical University; 3 The Graduate School, Shanxi University of Traditional Chinese Medicine
  • Received:2019-11-18 Revised:2020-05-17 Published:2020-08-15 Online:2020-08-12
  • Contact: △Corresponding Author E-mail: mych65@163.com E-mail:mych65@163.com

摘要: 目的 观察下调跨膜蛋白97(TMEM97)对卵巢癌细胞SKOV3增殖和凋亡的影响,并探讨其相关机制。方法 将卵巢癌细胞SKOV3分为转染组(si-TMEM97组)、阴性对照组(siNC组)和空白对照组(Control组)。采用实时荧光定量PCR(qPCR)及Western blot法分别在mRNA和蛋白表达水平评估转染siRNA-TMEM97后TMEM97的沉默效果;分别在转染后采用CCK-8法、PI染色法、Annexin V-FITC/PI染色法和流式细胞术检测下调TMEM97对细胞的增殖、细胞周期和细胞凋亡的影响,并检测B淋巴细胞瘤-2基因(Bcl-2)、Bcl-2相关X蛋白质(Bax)及P38丝裂原活化蛋白激酶(P38 MAPK)通路相关蛋白(P38/MAPK、p-P38/MAPK)的表达变化。结果 si-TMEM97组细胞的增殖能力相比Control组和siNC组降低,而早期凋亡率增高(均P<0.01)。si-TMEM97组细胞Bcl-2的蛋白相对表达量较Control组和siNC组降低,而Bax、p-P38/MAPK的蛋白相对表达量较Control组和siNC组增高(均P<0.05)。结论 下调TMEM97的表达使卵巢癌细胞SKOV3增殖减少,凋亡增加,这可能与调节凋亡相关蛋白Bcl-2/Bax表达及P38/MAPK信号通路的激活有关。

关键词: 卵巢肿瘤, 细胞增殖, 细胞凋亡, bcl-2相关X蛋白质, p38丝裂原活化蛋白激酶类, TMEM97, P38/MAPK信号通路

Abstract: Objective To explore the effects of transmembraneprotein 97 (TMEM97) on the proliferation and apoptosis of human ovarian cancer SKOV3 cells and its mechanism. Methods The SKOV3 cells were divided into transfection (si-TMEM97) group , negative control (siNC) group and blank control (Control) group. qPCR and Western blot assay were used to evaluate the TMEM97 silencing efficiency at mRNA and protein levels after siRNA-TMEM97 transfection. The proliferation ability of ovarian cancer cells was detected by CCK-8 assay after transfection. PI staining assay was used to observe the effect of down-regulated gene TMEM97 on cell cycle. The apoptosis rate was measured by Annexin V-FITC/PI assay. Western blot assay was used to evaluate the expressions of the B-cell lymphoma-2 (Bcl-2), Bcl-2 associated X (Bax) and P38 mitogen-activated protein kinase pathway-associated proteins (P38/MAPK and p-P38/MAPK). Results Compared with the Control group and the siNC group, the proliferation ability of SKOV3 cells was inhibited in the si-TMEM97 group, and the early apoptosis rate of SKOV3 cells was significantly increased (P<0.01). The relative expression of Bcl-2 protein in SKOV3 cells was significantly decreased, and the relative expressions of Bax protein and p-P38/MAPK were significantly increased in si-TMEM97 group than those of Control group and siNC group (all P<0.05). Conclusion The down-regulation of TMEM97 decreases the proliferation and increases the apoptosis of SKOV3 cells, which may be related with the regulating the Bcl-2/Bax expression and activating P38/MAPK signaling pathway.

Key words: ovarian neoplasms, cell proliferation, apoptosis, bcl-2-associated X protein, p38 mitogen-activated protein kinases, TMEM97, P38/MAPK signaling pathway

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