Hu Zhuoying, Zhang Hongyu, Tang Liangdan, Lou Meng, Geng Yanqing
Department of Obstetrics and Gynecology, The 1st Affiliated Hospital of Chongqing Medical University, Chongqing, China (mainland).
Department of General Surgery, The 1st Affiliated Hospital of Chongqing Medical University, Chongqing, China (mainland).
Med Sci Monit. 2017 Mar 16;23:1317-1324. doi: 10.12659/msm.900320.
BACKGROUND The role that nc886, a non-coding microRNA, plays in human endometrial cancer is unknown. The present study aimed to describe the functional role of nc886 in human endometrial cancer-1A (HEC-1A) cell line, which may provide another target for human endometrial cancer treatment. MATERIAL AND METHODS The expression levels of nv886 in normal human endometrial tissue and the early phase and late phase of human endometrial cancer tissues were determined and compared by fluorescence in situ hybridization (FISH). Small interference RNA (siRNA) was used to inhibit nc886, and cell proliferation was evaluated with the MTT test. mRNA levels of PKR, NF-κB, vascular endothelial growth factor (VEGF), and caspase-3 were determined against glyceraldehyde 3-phosphate dehydrogenase (GAPDH between the HEC-1A control group and the silenced group (nc886 silenced with siRNA) by real-time reverse transcription polymerase chain reaction (RT-PCR). The protein levels of PKR (total and phosphorylated form), NF-κB, VEGF, and caspase-3 were determined against GAPDH by Western blotting, and cell apoptosis was determined by flow cytometry. RESULTS Our results indicated that a higher level of nc886 was expressed in the late phase of human endometrial cancer tissue, less than in the early phase but still higher than in normal human endometrial tissue. After nc886 was silenced, protein levels of p-PKR (phosphorylated PKR) and caspase-3 were increased, whereas NF-κB and VEGF were decreased. CONCLUSIONS The rate of apoptosis in the silenced group was increased and the rate of cell proliferation was slower in comparison to the control.
背景 非编码微小RNA nc886在人类子宫内膜癌中所起的作用尚不清楚。本研究旨在描述nc886在人子宫内膜癌1A(HEC-1A)细胞系中的功能作用,这可能为人类子宫内膜癌治疗提供另一个靶点。
材料与方法 通过荧光原位杂交(FISH)检测并比较正常人类子宫内膜组织以及人类子宫内膜癌组织早期和晚期中nv886的表达水平。使用小干扰RNA(siRNA)抑制nc886,并通过MTT试验评估细胞增殖。通过实时逆转录聚合酶链反应(RT-PCR),以甘油醛-3-磷酸脱氢酶(GAPDH)为对照,测定HEC-1A对照组和沉默组(用siRNA沉默nc886)中PKR、NF-κB、血管内皮生长因子(VEGF)和半胱天冬酶-3的mRNA水平。通过蛋白质印迹法以GAPDH为对照测定PKR(总形式和磷酸化形式)、NF-κB、VEGF和半胱天冬酶-3的蛋白质水平,并通过流式细胞术测定细胞凋亡。
结果 我们的结果表明,nc886在人类子宫内膜癌组织晚期的表达水平较高,低于早期但仍高于正常人类子宫内膜组织。nc886沉默后,p-PKR(磷酸化PKR)和半胱天冬酶-3的蛋白质水平升高,而NF-κB和VEGF降低。
结论 与对照组相比,沉默组的细胞凋亡率增加,细胞增殖率较慢。