shRNA干扰LGR5对HeLa细胞增殖及耐药性的影响

目的探讨富含亮氨酸重复序列G-蛋白偶联受体5（LGR5）基因在HeLa细胞增殖及耐药性中的作用及可能机制。方法采用shRNA技术干扰LGR5表达,构建LGR5干扰（shLGR5）的HeLa细胞,并设对照组（shCon）。采用细胞计数、平板克隆以及MTT实验观察shLGR5对HeLa细胞增殖及耐药性的影响;采用Western blot检测LGR5、β-catenin在HeLa细胞中的表达。结果成功构建了LGR5干扰的HeLa细胞系;与shCon组相比,shLGR5组细胞生长速度及克隆形成率明显降低,差异具有统计学意义（P〈0.01）;shLGR5组与shCon组HeLa细胞对顺铂的耐药性差异亦具...

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Published in西安交通大学学报（医学版） Vol. 38; no. 2; pp. 206 - 209
Main Author 陈庆 赵金燕 卢晓宁 郑鹏生
Format Journal Article
LanguageChinese
Published 西安交通大学第二附属医院妇产科,陕西西安,710004%西安交通大学第一附属医院生殖医学科,陕西西安,710069 2017
Subjects
blot
HeLa细胞
LGR5
shRNA
Western
β-catenin
细胞增殖
耐药性
Western blot
β-catenin
Westernblot
cell proliferation
shRNA
HeLa cell
耐药性
HeLa细胞
drug resistance
LGR5
细胞增殖
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ISSN1671-8259
DOI10.7652/jdyxb201702010

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Summary:目的探讨富含亮氨酸重复序列G-蛋白偶联受体5（LGR5）基因在HeLa细胞增殖及耐药性中的作用及可能机制。方法采用shRNA技术干扰LGR5表达,构建LGR5干扰（shLGR5）的HeLa细胞,并设对照组（shCon）。采用细胞计数、平板克隆以及MTT实验观察shLGR5对HeLa细胞增殖及耐药性的影响;采用Western blot检测LGR5、β-catenin在HeLa细胞中的表达。结果成功构建了LGR5干扰的HeLa细胞系;与shCon组相比,shLGR5组细胞生长速度及克隆形成率明显降低,差异具有统计学意义（P〈0.01）;shLGR5组与shCon组HeLa细胞对顺铂的耐药性差异亦具有统计学意义（P〈0.01）;且干扰LGR5抑制HeLa细胞中β-catenin蛋白的表达。结论 LGR5基因在HeLa细胞增殖及耐药性中有重要作用,且与Wnt/β-catenin通路有关。
Bibliography:Objective To investigate the effect of leucine-rich-repeat-containing G-protein-coupled receptor 5（LGR5）on the proliferation and drug resistance of HeLa cells and its possible mechanism.Methods LGR5 expression was interfered using shRNA,and LGR5 knockdown HeLa cells were constructed.The effect of LGR5 on the proliferation and drug resistance of HeLa cell was evaluated by cell count,clone formation and MTT;the expressions of LGR5 andβ-catenin in HeLa cells were detected by Western blot method.Results LGR5 knockdown HeLa cell line was successfully constructed;the cell growth rate and clone formation rate in shLGR5 group were markedly decreased compared to those in shCon group（P〈0.01）.Drug resistance of HeLa to cisplatin differed significantly between shLGR5 group and shCon group（P 〈0.01）.Moreover,the LGR5 knockdown inhibited the expression ofβ-catenin in HeLa cells.Conclusion LGR5 plays an important role in cell proliferation and drug resistance of HeLa cells,and its mechanism is related to Wnt/β-catenin signal
ISSN:1671-8259
DOI:10.7652/jdyxb201702010