PERK在肝癌中的表达及其对肝癌细胞侵袭和索拉菲尼抵抗的影响

Expression of PERK in hepatocellular carcinoma and its effect on invasion of hepatocellular carcinoma cells and sorafenib resistance

目的观察蛋白激酶RNA样内质网激酶(PERK)在肝细胞癌和对应癌旁组织中的表达,探究PERK基因在肝癌细胞侵袭和索拉菲尼抵抗中的作用。方法免疫组化法检测PERK在人肝癌组织中的表达水平并分析其表达差异与临床病理特征之间的关联性;利用脂质体转染干扰序列至肝癌细胞,分为3组:对照组、sPNC转染组、P-/ERK转染组,Western blot分析HepG2细胞基质金属蛋白酶T(MMP2)和基质金属蛋白酶T(MMP9)的表达变化,Tonsweli小室法检查肝癌细胞的侵袭性能;HepG2细胞转染干扰序列后作用于索拉菲尼,分为4组:对照组、索拉菲尼组、索拉菲尼+w-NC转染组、索拉菲尼+isi-/ERK转染组,流式细胞术分析各组凋亡细胞比率。结果相比于癌旁组织,PERK在肝癌组织中明显高表达,同时PERK高表达与肿瘤的血管侵犯具有关联性;相比于对照组和P-NC转染组,sPPERK转染组MMP2和MMP9的蛋白表达明显降低、侵袭的细胞数目减少;PERK的沉默显著提高HepG2细胞在索拉菲尼诱导下发生的凋亡。结论PERK蛋白在肝癌组织中高表达且与肿瘤的侵袭能力和索拉菲尼抵抗密切相关.

Objective To observe the expression of protein kinase RNA-like endoplasmic reticulum kinase(PERK)in the hepatocellular carcinoma tissues and adjacent tissues,and to explore the role of PERK in the invasion of liver cancer cells and sorafenib resistance.Methods Immumnohistochemistry was used to detect the expression of PERK in human liver cancer tissues,the correlation between PERK expression differences and clinicopathological features was analyzed.The interference sequence was transfected with lipofectine to HepG2 cells,divided into 3 groups,control group,si-NC transfection group and si-PERK transfection group,and the expression of matrix metalloprotei-nase-2(MMP2)and matrix metalloproteinase-9(MMP9)were detected by Western blot,invasion ability of liver cancer cells was detected by transwell matrigel invasion assay.HepG2 cells was treated with sorafenib after trans-fecting with siRNA and divided into 4 groups:control group,sorafenib group,sorafenib+si-NC transfection group,sorafenib+si-PERK transfection group,proportion of apoptotic cells was detected by flow cytometry.Results PERK was significantly higher expressed in liver cancer tissues than in adjacent tsses and the expression of PERK was associated with tumor vascular invasion.Compared with the groups of control and transfection/Si-NC,the ex-pression of MMP2 and MMP9 was signifcantly decreased and the number of invaded cells was obviously reduced in the group of transfection/si-PERK.PERK silencing significantly promoted the apoptosis of HepG2 cells induced by sorafenib.Conclusion PERK is highly expressed in hepatoellular carcinoma tissues and is closely related to tumor invasion and sorafenib resistance.