天津医药

天津医药 ›› 2020, Vol. 48 ›› Issue (6): 512-516.

• 细胞与分子生物学 • 上一篇    下一篇

lncRNA HOST2通过PI3K/Akt途径调控卵巢癌SKOV3 细胞对顺铂的敏感性

韩建秋 1，郝丽惠 1，邢雅玲 1，张艳 1，吴维光 2△
  

  1. 1 天津市泰达医院妇科（邮编 300457）；2 中国人民武装警察部队特色医学中心妇产科

  • 收稿日期:2019-08-26 修回日期:2020-04-23 出版日期:2020-06-15 发布日期:2020-06-15
  • 通讯作者: 吴维光 E-mail:doctoratewu@126.com
  • 作者简介:作者晋升，7月前

Long noncoding RNA HOST2 regulates the sensitivity of human ovarian carcinoma SKOV3 cells to cisplatin through PI3K/Akt pathway

HAN Jian-qiu1, HAO Li-hui1, XING Ya-ling1, ZHANG Yan1, WU Wei-guang2△ #br#   

  1. 1 Department of Gynecology, TEDA Hospital, Tianjin 300457, China; 2 Department of Obstetrics and Gynecology, the
    Characteristic Medicine Center of the Chinese People    ’s Armed Forces
  • Received:2019-08-26 Revised:2020-04-23 Published:2020-06-15 Online:2020-06-15

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摘要： 摘要：目的 探讨下调长链非编码RNA（lncRNA）上皮性卵巢癌转录因子2（HOST2）表达对人卵巢癌SKOV3细 胞顺铂敏感性的影响及其作用机制。方法 体外培养人卵巢癌SKOV3细胞，取对数生长期细胞分为空白对照组、阴 性对照组和siRNA干扰组，其中阴性对照组和siRNA干扰组SKOV3细胞应用Lipofectamine 2000分别转染阴性对照 siRNA 和 lncRNA HOST2-siRNA，空白对照组未进行转染。采用 qPCR 法检测各组细胞 lncRNA HOST2 的表达； Western blot检测磷脂酰肌醇3-激酶（PI3K）/蛋白质丝氨酸苏氨酸激酶（Akt）信号通路相关蛋白Akt、p-Akt （S473）、pAkt（S380）及Bcl-2的表达；CCK-8法检测经不同浓度（0、20、40、60、80、100 μmol/L）顺铂作用后细胞的存活情况，并计算半抑制浓度（IC50）；流式细胞术检测经20 μmol/L顺铂作用后的细胞凋亡率。结果 与空白对照组和阴性对照组 比较，siRNA 干扰组细胞 lncRNA HOST2 表达及 p-Akt（S473）、p-Akt（S380）和 Bcl-2 蛋白表达水平均降低（均 P＜ 0.05）；3组Akt蛋白表达水平差异无统计学意义（P＞0.05）。空白对照组、阴性对照组和siRNA干扰组细胞存活率均 随顺铂药物浓度的增加而降低，IC50分别为（59.58±5.97）、（51.42±5.22）和（39.75±5.31）μmol/L。siRNA干扰组细胞凋 亡率（12.42%±1.46%）明显高于空白对照组（7.53%±1.25%）和阴性对照组（8.16%±1.31%）。结论 下调 lncRNA HOST2表达可增强人卵巢癌SKOV3细胞对顺铂的敏感性，其机制与抑制PI3K/Akt信号通路相关蛋白的表达有关

关键词: 卵巢肿瘤, RNA, 长链非编码, 上皮性卵巢癌转录因子2, 顺铂, PI3K/Akt途径

Abstract: Abstract: Objective To investigate the effect of down-regulation of long noncoding RNA (lncRNA) human ovarian cancer-specific transcripts 2 (HOST2) expression on the sensitivity of human ovarian carcinoma SKOV3 cells to cisplatin, and its possible mechanism thereof. Methods SKOV3 cells were cultured in vitro, and the logarithmic growth phase cells were divided into blank control (BC) group, negative control (NC) group and siRNA group. The siRNAs targeting lncRNA HOST2 and negative control were transfected into SKOV3 cells of siRNA group and NC group by cationic liposome, the BC group was not transfected. The expression of lncRNA HOST2 was detected by qPCR, and the protein expressions of Akt, pAkt (S473), p-Akt (S380) and Bcl-2 were detected by Western blot assay. The cell survival was detected by CCK-8 after SKOV3 cells were treated by different concentrations of cisplatin (0、20、40、60、80、100 μmol/L) and the cisplatin half inhibitory concentration (IC50) was calculated. The apoptotic rate after treatment with 20 μmol/L cisplatin was checked by flow cytometry. Results Compared with BC group and NC group, the expression of lncRNA HOST2 and the protein expressions of p-Akt (S473), p-Akt (S380) and Blc-2 decreased in siRNA group (P＜0.05). There was no significant difference in the expression of Akt protein between three groups (P＞0.05). The cell survival rates all decreased with the increased cisplatin concentration, and values of IC50 were (59.58±5.97), (51.42±5.22) and (39.75±5.31)μmol/L in BC group, NC group and siRNA group. The apoptotic rate was significantly higher in the siRNA group (12.42%±1.46%) than that in the BC group (7.53%±1.25%) and the NC group (8.16%±1.31%). Conclusion The downregulation of lncRNA HOST2 expression could enhance the cisplatin sensitivity in human ovarian cancer SKOV3 cells, and its mechanism is related to the inhibition of PI3K/Akt pathway-related protein expressions.

Key words: ovarian neoplasms, RNA, long noncoding, ovarian cancer-specific transcripts 2, cisplatin, PI3K/Akt
pathway