摘要
【目的】基于FAK/PI3K/Akt通路,探讨FAK抑制剂CT-707对人肝癌HCC-LM3细胞骨架重排、侵袭、迁移及裸鼠皮下移植瘤生长的影响和机制。【方法】HCC-LM3细胞分为Control组、CT-707低剂量(1.5μmol/L)组、CT-707中剂量(3μmol/L)组、CT-707高剂量(6μmol/L)组,采用Transwell小室实验、细胞划痕、MTT实验分别测定细胞侵袭、迁移能力及细胞活力;采用RT-qPCR和Western blot分别检测Palladin、Vimentin、MMP2和MMP9 mRNA及蛋白的表达;Western blot检测p-FAK、FAK、p-PI3K、PI3K、p-Akt及Akt蛋白的表达。使用HCC-LM3细胞系建立裸鼠皮下移植瘤模型,分为模型组、CT-707组(20 mg/kg,腹腔注射),每组6只,记录肿瘤体积及质量,采用HE染色法观察移植瘤组织结构变化,免疫组化检测移植瘤FAK、PI3K、p-Akt、MMP-2、MMP-9的表达。【结果】与Control组比较,CT-707高、中、低剂量组可显著抑制HCC-LM3细胞侵袭迁移能力,降低细胞活力,下调Palladin、Vimentin、MMP2、MMP9、p-FAK/FAK、p-PI3K/PI3K、p-Akt/Akt的表达水平(P<0.05),中、高剂量组的效应明显优于低剂量组(P<0.05),中、高剂量组间结果差异无统计学意义(P>0.05)。与模型组比较,CT-707治疗后可显著降低移植瘤体积和质量以及FAK、PI3K、p-Akt、MMP-2、MMP-9蛋白表达(P<0.05),抑瘤率为51.92%。【结论】CT-707可能通过抑制FAK/PI3K/Akt信号通路活性,降低细胞骨架重排及基质金属蛋白酶分泌,从而抑制肝癌细胞侵袭、迁移以及肿瘤生长。
【Objective】To explore the effect of FAK inhibitor CT-707 on the cytoskeleton rearrangement,invasion and migration of human hepatocellular carcinoma cells HCC-LM3 and its mechanism,and the growth of subcutaneous xenografted tumors in nude mice based on the FAK/PI3K/Akt pathway.【Methods】HCC-LM3 cells were divided into Control group,CT-707 low dose(1.5μmol/L)group,CT-707 medium dose(3μmol/L)group,CT-707 high dose(6μmol/L)group.Transwell chamber test,cell scratch,and MTT test were used to determine cell invasion,migration and cell viability.RT-qPCR and Western blot were used to detect the mRNA and protein expression of Palladin,Vimentin,MMP2 and MMP9.Western blot was used to detect the expression of p-FAK,FAK,p-PI3K,PI3K,p-Akt and Akt protein.HCC-LM3 cell line was used to establish subcutaneous xenograft tumor models in nude mice,which were divided into model group and CT-707 group(20 mg/kg,ip),with 6 mice in each group.The tumor volume and mass were recorded,the tissue structure changes of the transplanted tumor were observed by HE staining method,and the expression of FAK,PI3K,p-Akt,MMP-2,MMP-9 of the transplanted tumor was detected by immunohistochemistry.【Results】Compared with the control group,the CT-707 high,medium and low dose groups could significantly inhibit the invasion and migration ability,decrease cell viability,significantly down-regulate the expression levels of Palladin,Vimentin,MMP2,MMP9,p-FAK/FAK,p-PI3K/PI3K and p-Akt/Akt,(P<0.05),and the effect of the middle and high-dose groups was significantly better than that of the low-dose group(P<0.05),and there was no significant difference in the results between the middle and high-dose groups(P>0.05).Compared with those in the control group,CT-707 could significantly reduce the volume and quality of transplanted tumors and the protein expression of FAK,PI3K,p-Akt,MMP-2,and MMP-9 in the treatment groups(P<0.05),and the tumor inhibition rate was 51.92%.【Conclusion】CT-707 may inhibit the invasion and migration of hepatocellular carcinoma cells by inhibiting the activity of FAK/PI3K/Akt signaling pathway,thereby inhibiting cytoskeletal rearrangement and matrix metalloproteinase secretion.
作者
姚红兵
肖芳
郭威
吴嘉兴
文雪霖
蒋建晖
华赟鹏
YAO Hong-bing;XIAO Fang;GUO Wei;WU Jia-xing;WEN Xue-lin;JIANG Jian-hui;HUA Yun-peng(Department of Hepatobiliary Surgery,The Second Affiliated Hospital of Guilin Medical University,Guilin 541199,China;Department of Oncology,The 924th Hospital of PLA Joint Service Support Force Liuzhou Worker's Hospital,Guilin 541002,China;Department of Liver surgery,The First Affiliated Hospital of Sun Yat-sen University,Guangzhou 510080,China)
出处
《中山大学学报（医学科学版）》
CAS
CSCD
北大核心
2021年第3期364-372,共9页
Journal of Sun Yat-Sen University:Medical Sciences
基金
广西自然科学基金(2018JJA140512)。
关键词
CT-707
FAK
肝癌
侵袭
迁移
CT-707
focaladhesionkinase
hepatic carcinoma
invasion,migration
