下调跨膜蛋白16A对子宫肉瘤FU-MMT-1细胞增殖侵袭的影响及初探其机制

Effects of down-regulation of transmembrane protein 16A on the proliferation and invasion of uterine carcinosarcomaa FU-MMT-1 cells and a preliminary investigation of its mechanism

目的 通过研究下调跨膜蛋白16A(TMEM16A)对子宫肉瘤FU-MMT-1细胞增殖侵袭及患者生存率的影响,初步探究其抑制子宫肉瘤的作用机制。方法 将子宫肉瘤FU-MMT-1细胞分为对照组和敲减组。用细胞计数试剂盒-8法检测子宫肉瘤FU-MMT-1细胞增殖情况,用Transwell实验检测细胞侵袭能力。在The Cancer Genome Atlas(TCGA)大数据库中下载子宫肉瘤患者的基因数据,根据TMEM16A和转录因子κB(NF-κB)表达的中位值将患者分为高表达组和低表达组进行生存分析,用蛋白质印迹(Western blot)法检测细胞TMEM16A和p65蛋白的表达情况。结果 子宫肉瘤FU-MMT-1细胞对照组和敲减组的细胞增殖率分别为(100.00±0)%和(62.17±5.40)%;侵袭细胞数分别为(255±23)和(80±6)个。Kaplan-Meier生存分析显示:TMEM16A高表达子宫肉瘤患者预后生存期显著降低;NF-κB高表达子宫肉瘤患者预后生存期显著降低,TMEM16A与NF-κB共同高表达组总生存显著低于共同低表达组,上述指标差异均有统计学意义(均P<0.05);对照组和敲减组FU-MMT-1细胞核中p65蛋白相对表达水平分别为0.88±0.03和0.21±0.10,差异有统计学意义(P<0.05),细胞质中p65蛋白表达水平分别为0.40±0.05和0.80±0.05,差异有统计学意义(P<0.05)。结论 下调TMEM16A影响p65蛋白表达抑制子宫肉瘤FU-MMT-1细胞的增殖和侵袭。

Objective To investigate the effect of down-regulation of transmembrane protein 16A(TMEM16A)on the proliferation and invasion of uterine carcinosarcoma FU-MMT-1 cells and the overall survival rate of patients,and to preliminarily explore its mechanism.Methods Uterine carcinosarcoma FU-MMT-1 cells were divided into control group and knockdown group.Uterine carcinosarcoma FU-MMT-1 cell proliferation was detected by applying the cell counting Kit-8 method;cell invasion ability was detected by applying the Transwell assay;gene transcription data information of uterine carcinosarcoma patients was downloaded from The Cancer Genome Atlas(TCGA)database,and patients were divided into high and low expression groups according to the median values of TMEM16A and transcription factor-κB(nuclear factor-κB)expression.Overall survival rates were analyzed.Western blot was applied to detect the expressions of cellular TMEM16A and p65 protein.Results The cell proliferation rates of the control and knockdown groups of uterine carcinosarcoma FU-MMT-1 cells were(100.00±0)%and(62.17±5.40)%respectively;the number of invasive cells was(255±23)and(80±6)respectively;Kaplan-Meier survival analysis showed that the overall survival of uterine carcinosarcoma patients with TMEM16A high expression was significantly reduced;the overall survival with NF-κB high expressionwas significantly reduced,and the overall survival of the TMEM16A and NF-κB co-high expression groupwas significantly lower than that of co-low expression group;the differences in the above indicators were statistically significant(all P<0.05);the relative expression levels of p65 protein in nucleus of FU-MMT-1 cells in the control and knockdown groups were 0.88±0.03 and 0.21±0.10 with statistically significant differences(P<0.05);p65 protein expression levels in cytosol were 0.40±0.05 and 0.80±0.05 with statistically significant differences(P<0.05).Conclusion Down-regulation of TMEM16A can activate NF-κB signaling pathway and inhibit proliferation and invasion of Uterine Carcinosarcoma FU-MMT-1 cells.