缝隙连接蛋白26基因修饰对人高转移性肝癌细胞增殖和侵袭能力的影响

发布时间：2018-06-28 07:01

本文选题：缝隙连接蛋白 + 肝癌细胞　；参考：《中国老年学杂志》2017年02期

【摘要】：目的探讨缝隙连接蛋白(Cx)26基因对人高转移性肝癌细胞株(HCCLM3)细胞增殖和侵袭能力的影响。方法采用慢病毒载体(LV)转染法将载体LV-Cx26转染高转移性HCCLM3细胞为研究组,仅转染空载体LV-NC-Cx26为对照组,未转染的HCCLM3细胞为空白组。RTPCR和Western印迹测定转染后Cx26 mRNA和蛋白表达;划痕荷载染料传输实验检测细胞通讯功能;流式细胞术检测细胞周期并通过CCK-8增殖实验、Transwell细胞侵袭实验检测细胞增殖、侵袭能力。结果研究组Cx26 mRNA和蛋白表达量均明显高于对照组和空白组(P0.05)。研究组转染慢病毒载体LV-Cx26后的细胞通讯功能明显强于对照组、空白组,即划痕细胞中的荧光染料可传递到相邻的3~4列细胞。研究组G0/G1期细胞数明显多于对照组和空白组,S期细胞数明显少于对照组和空白组(P0.05)。M期三组细胞数无统计学差异(P0.05)。CCK-8法检测显示,72 h起研究组细胞增殖速度明显低于同期对照组和空白组(P0.05);对照组和空白组各时间点细胞增殖情况无统计学差异(P0.05)。Transwell细胞侵袭实验显示,研究组穿膜细胞数明显少于对照组和空白组(P0.05)。结论转染Cx26基因可有效抑制HCCLM3细胞的增殖和侵袭能力,降低恶性生物学特征,可成为晚期肝癌治疗的新思路。
[Abstract]:Objective to investigate the effect of gap junction protein (CX) 26 gene on proliferation and invasion of human hepatocellular carcinoma cell line (HCCLM3). Methods LV-Cx26 was transfected into high metastatic HCCLM3 cells by using lentivirus vector (LV) transfection method, only the empty vector LV-NC-Cx26 was used as control group. The untransfected HCCLM3 cells were treated with blank group. RT-PCR and Western blot were used to detect the expression of Cx26 mRNA and protein. The cell cycle was detected by flow cytometry and the cell proliferation and invasiveness were detected by Transwell cell invasion assay with CCK-8 proliferation assay. Results the expression of Cx26 mRNA and protein in study group was significantly higher than that in control group and blank group (P0.05). The cell communication function of lentivirus vector LV-Cx26 was stronger than that of control group. The fluorescent dye from scratch cells in blank group could be transferred to the adjacent 3Cx26 cells. The number of cells in the G _ 0 / G _ 1 phase in the study group was significantly higher than that in the control group and the blank group. The number of cells in the S phase of the study group was significantly lower than that in the control group and the blank group (P0.05) .There was no statistical difference among the three groups (P0.05). At the same time, the control group and the blank group (P0.05); the control group and the blank group at each time point cell proliferation no statistical difference (P0.05) .Transwell cell invasion test showed, The number of perforating cells in the study group was significantly less than that in the control group and the blank group (P0.05). Conclusion transfection of Cx26 gene can effectively inhibit the proliferation and invasion of HCCLM3 cells, reduce the malignant biological characteristics, and become a new approach for the treatment of advanced liver cancer.
【作者单位】：暨南大学附属第一临床学院华侨医院胃肠外科;
【基金】：广东省科学技术厅计划项目(No.2016ZC0142)
【分类号】：R735.7

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