沉默CXCR4基因对人子宫颈癌HeLa细胞增殖、侵袭及上皮间质转化相关蛋白表达的影响  

Effect of silencing CXCR4 gene on proliferation, invasion and epithelial-mesenchymal transition related protein expressions in HeLa cells of human cervical cancer

作　　者：李风艳[1] 于利君[1] 张三元[1] 李莉[1] 张燕[1] Li Fengyan, Yu Lijun, Zhang Sanyuan, Li Li, Zhang Yon(Department of Gynecology, the First Hospital of Shanxi Medical University, Taiyuan 030001, Chin)

机构地区：[1]山西医科大学第一医院妇科,太原030001

出　　处：《肿瘤研究与临床》2018年第3期152-156,共5页Cancer Research and Clinic

基　　金：山西省基础研究计划（2015011112）

摘　　要：目的探讨小分子干扰RNA（siRNA）干扰CXCR4基因对人子宫颈癌HeLa细胞增殖、侵袭及上皮间质转化因子Twist、Slug及E钙黏蛋白（E-cad）表达的影响。方法体外培养HeLa细胞，设计合成CXCR4基因siRNA（CXCR4-siRNA）转染至HeLa细胞，采用四甲基偶氮唑盐（MTT）法检测细胞增殖能力的变化，Transwell实验观察细胞侵袭能力的变化，蛋白印迹法检测CXCR4、Twist、Slug、E-cad蛋白表达的变化。结果与空白组及阴性对照组相比，CXCR4-siRNA转染至HeLa细胞后能抑制CXCR4蛋白的表达（0.959±0.197、0.932±0.141和0.379±0.022，F=16.286，P=0.004）。MTT结果显示：48及72 h时，CXCR4-siRNA组HeLa细胞的增殖活性受到抑制（48 h：0.846±0.034、0.823±0.025和0.744±0.039，F=7.379，P=0.024；72 h：0.996±0.026、0.964±0.059和0.829±0.051，F=10.425，P=0.011）。Transwell结果显示：48及72 h时，CXCR4-siRNA组HeLa细胞的侵袭力受到抑制（细胞穿膜数量为48 h：37.4±8.1、33.6±6.2和25.4±3.2，F=4.830，P=0.029；72 h：48.4±7.6、43.0±5.4和33.4±6.7，F=6.579，P=0.012）。转染后48 h，CXCR4-siRNA组HeLa细胞Twist及Slug蛋白的表达降低（Twist：0.93±0.11、1.00±0.17和0.53±0.07，F=10.962，P=0.010；Slug：0.52±0.08、0.47±0.06和0.19±0.03，F=25.579，P=0.001），E-cad的表达增高（0.53±0.04、0.55±0.24和1.11±0.14，F=12.494，P=0.007）。结论沉默CXCR4基因的表达可以抑制HeLa细胞增殖，降低其侵袭力，通过下调Twist、Slug蛋白的表达，上调E-cad表达，抑制HeLa细胞上皮间质转化。ObjectiveTo investigate the effects of small interfering RNA （siRNA） CXCR4 gene on proliferation, invasion and epithelial-mesenchymal transition factors including Twist, Slug and E-cadherin （E-cad） in HeLa cells of human cervical carcinoma.MethodsHeLa cells were cultured in vitro and CXCR4-siRNA was transfected into HeLa cells. Methyl thiazolyl tetrazolium （MTT） assay was used to detect the changes of cell proliferation. Transwell experiment was used to observe the changes of cell invasion, and the expressions of CXCR4, Twist, Slug and E-cad protein were detected by Western blot.ResultsCompared with the blank control group and the negative control group, CXCR4-siRNA transfected into HeLa cells significantly inhibited the expression of CXCR4 protein （0.959±0.197, 0.932±0.141 vs. 0.379±0.022 respectively; F=16.286, P=0.004）. MTT results showed that the proliferation of HeLa cells was inhibited at 48 h and 72 h （48 h： 0.846 ± 0.034, 0.823 ± 0.025 vs. 0.744 ± 0.039, F= 7.379, P= 0.024; 72 h： 0.996±0.026, 0.964±0.059 vs. 0.829±0.051, F= 10.425, P= 0.011）. Transwell results showed that the invasiveness of HeLa cells in CXCR4-siRNA group was inhibited at 48 h and 72 h （cell membrane number, 48 h： 37.4±8.1, 33.6±6.2 vs. 25.4±3.2, F = 4.830, P = 0.029. 72 h： 48.4±7.6, 43.0±5.4 vs. 33.4±6.7, F = 6.579, P = 0.012）. After transfection for 48 h, the expression of Twist and Slug protein was decreased and E-cad protein was increased in CXCR4-siRNA group （Twist： 0.93±0.11, 1.00±0.17 vs. 0.53±0.07, F = 10.962, P = 0.010; Slug： 0.515±0.084, 0.470±0.055 vs. 0.190±0.028, F = 25.579, P = 0.001; E-cad： 0.53±0.04, 0.55±0.24 vs. 1.11±0.14, F = 12.494, P = 0.007）.ConclusionsSilencing CXCR4 gene can inhibit the proliferation and reduce the invasiveness of HeLa cells. Inhibiting EMT of HeLa cells can be achieved by down-regulating the expressions of Twist and Slug protein, and up-regulating the expression of E-cad.

关 键 词：宫颈肿瘤 CXCR4 上皮间质转化 

分 类 号：R737.33[医药卫生—肿瘤]