摘要
目的 探讨Krǜppel样因子4(KLF4)对胰腺癌上皮间质转化(EMT)和侵袭能力的影响.方法 采用免疫组织化学染色法检测KLF4在70例胰腺癌和10例胰腺正常组织中的表达情况,分析其与临床病理特征的相关性.构建针对KLF4的小发卡RNA(shKLF4)和阴性对照shRNA,分别转染胰腺癌PANC1细胞,采用实时荧光定量PCR和蛋白质印迹法检测KLF4、E-钙黏素(E-cadherin)、波形蛋白(vimentin) mRNA及蛋白的表达;采用划痕实验、Transwell小室检测细胞的侵袭和转移能力.结果 KLF4在胰腺癌组织中的阳性表达率(47.1%)显著低于胰腺正常组织(80.0%),且KLF4表达与胰腺癌肿瘤分级、分期和远处转移呈负相关.降低PANC1细胞中KLF4的表达后,E-cadherin的mRNA和蛋白表达水平均明显下调(F=25.71,P=0.0011),vimentin的mRNA和蛋白表达水平均明显上调(F =24.95,P=0.0012),细胞形态由上皮表型向间质表型转化,细胞划痕愈合能力(F=47.82,P<0.001)、细胞迁移(F=53.68,P=0.0001)和侵袭(F=27.65,P=0.0009)能力均显著增强.结论 抑制KLF4表达可促进胰腺癌细胞EMT,并增强侵袭能力.
Objective To explore the effect of Kruppel like factor 4 (KLF4) on epithelial-to-mesenchymal transition and invasion ability of pancreatic cancer.Methods The expression of KLF4 in 70 pancreatic cancer tissues and 10 normal pancreatic tissues was detected by immunohistochemistry,and the correlations between KLF4 expression and pathological characteristics were analyzed.Small hairpin RNA targeting KLF4 (sh-KLF4) and negative control shRNA were constructed.After the transfection of shRNA,qRT-PCR and western blot were used to detect the mRNA and protein expression of KLF4,E-cadherin and vimentin,and cell scratch-wound assay and transwell assay were utilized to determine the ability of invasion and metastasis.Results KLF4 expression (47.1%) was lower in pancreatic cancer tissues compared with normal pancreatic tissues (80.0%),and negatively correlated with cell differentiation,tumor stage and distant metastasis.Down-regulated KLF4 expression in PANC1 cell caused decreased mRNA and protein expression of E-cadherin (F =25.71,P =0.0011) and increased mRNA and protein expression of vimentin (F =24.95,P=0.0012).Knockdown of KLF4 in PANC1 cell promoted the transition from epithelial morphology to mesenchymal morphology,and enhanced the healing ability (F =47.82,P < 0.001),migration (F =53.68,P=0.0001) and invasion (F=27.65,P=0.0009).Conclusions Knockdown of KLF4 can promote EMT and enhance the invasion ability of pancreatic cancer.
作者
徐向辉
赵忠
赵国栋
盛金鑫
Xu Xianghui;Zhao Zhong;Zhao Guodong;Sheng Jinxin(Department of General Surgery,Haimen People's Hospital,Haimen 226100,China)
出处
《中华胰腺病杂志》
CAS
2019年第1期38-42,共5页
Chinese Journal of Pancreatology
