摘要
目的 观察shRNA Twist基因对人绒毛膜癌细胞JEG-3增殖、侵袭和凋亡的影响。方法 以人源性绒毛膜癌细胞为种子细胞,构建shRNA-Twist小干扰RNA载体,通过慢病毒转染绒毛膜癌细胞,根据转染慢病毒质粒的不同分成3组,即空白对照组、空白质粒组和shRNA干扰组。RT-qPCR和Western Blot检测Twist和凋亡蛋白caspase-3和caspase-9的表达。Transwell法检测人绒毛膜癌细胞侵袭能力,CCK-8法检测人绒毛膜癌细胞增殖能力,AV-PI试剂盒检测人绒毛膜癌细胞凋亡水平。结果 RT-PCR结果显示,空白质粒组的Twist的mRNA的相对表达量(与空白对照组相比)为(1.011±0.125);shRNA干扰组的Twist的mRNA的相对表达量(与空白对照组相比)为(0.377±0.021),高于空白质粒组的mRNA表达水平(P < 0.05)。Caspase-3和Caspase-9的mRNA含量具有同样的趋势。Western-Blot结果显示,空白质粒组和shRNA干扰组的的Twist蛋白相对表达量(与空白对照组相比)分别为(7.211±0.934)和(0.213±0.091),2组比较差异具有统计学意义(P < 0.05)。Caspase-3和Caspase-9的蛋白含量具有同样的趋势。空白对照组、空白质粒组和shRNA干扰组的Transwell结果比较具有明显的统计学差异(F = 23.983,P < 0.05)。shRNA干扰组与空白质粒组的Transwell结果比较具有明显的统计学差异(q =10.322,P < 0.05)。CCK-8和AV-PI检测结果具有同样的趋势。结论 shRNA-Twist可以明显的抑制绒毛膜癌细胞的增殖和侵袭,促进细胞凋亡,可以作为绒毛膜癌新的治疗靶基因。
Objective To explore the effects of shRNA Twist gene on JEG-3 proliferation, invasion and apoptosis in human choriocarcinoma cells. Methods Using human choriocarcinoma cells as seed cells, shRNA-Twist small interfering RNA vectors were constructed and transfected into choriocarcinoma cells by lentiviruses. The cells were divided into three groups according to the different lentiviral plasmids transfected, namely blank control group, blank plasmid group and shRNA interference group. RT-q PCR and Western Blot were used to detect the expressions of Twist and Caspase-3 and caspase-9. Transwell assay was used to detect the invasive ability of human choriocarcinoma cells,CCK-8 assay was used to detect the proliferation ability of human choriocarcinoma cells, and AV-PI kit was used to detect the apoptotic level of human choriocarcinoma cells. Results RT-PCR results showed that the relative expression of Twist in the blank plasmid group (compared with the blank control group) was (1.011 ± 0.125);the relative expression of Twist in the shRNA interference group (compared with the blank control group) was (0.377 ± 0.021), which was higher than that in the blank plasmid group (P < 0.05). The contents of Caspase-3 and Caspase-9 had the same trend. Western-Blot results showed that the relative expressions of Twist protein in the blank plasmid group and shRNA interference group (compared with the blank control group) were (7.211 ±0.934) and (0.213 ±0.091), respectively, with significant difference between the two groups (P < 0.05). The protein contents of Caspase-3 and Caspase-9 showed the same trend. Transwell results of blank control group, blank plasmid group and shRNA interference group were significantly different (F =23.983,P < 0.05). There were significant statistical differences in Transwell results between shRNA interference group and blank plasmid group (q = 10.322, P = 0.0157 < 0.05). CCK-8 and AV-PI test results showed the same trend. Conclusions shRNA-Twist can significantly inhibit the proliferation and invasi
作者
谢环
田洁
黄娅芬
XIE Huan;TIAN Jie;HUANG Ya-fen(Dept. of Gynecology,The 2nd People's Hospital of Yichang ,The Second People's Hospital of China Three Gorges University Gynecology,Yichang Hubei 443000;Dept. of Gynecology,Maternal and Child Health Hospital,Hubei Eastern Medical Group,Huangshi Hubei 435000,China)
出处
《昆明医科大学学报》
CAS
2019年第5期18-23,共6页
Journal of Kunming Medical University
基金
湖北省自然科学基金资助项目(2015CFB574)
