RIP1在肝癌组织中的表达意义及其对细胞侵袭、迁移的影响
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摘要
探讨受体相互作用蛋白激酶-1(RIP1)在肝癌组织及癌旁组织中的表达情况,并探索其对肝癌细胞侵袭和迁移的分子调控机制。收集肝癌和癌旁组织,免疫组化检测RIP1蛋白的表达,筛选高表达RIP1的肝癌细胞系。对肝癌细胞系进行RIP1沉默处理并设置阴性对照。采用western blot及RT-PCR法检测RIP1的表达量;Tranwell观察细胞侵袭能力;细胞划痕观察细胞迁移能力。结果显示,48例肝癌组织中,22例存在RIP1蛋白阳性表达,阳性率为45.83%,其中,强阳性患者为15例,中等及弱阳性7例,该研究中有6例复发患者。RIP1在肝癌组织中的表达明显高于癌旁组织(P<0.05),高表达RIP1患者的总生存率和无复发生存率显著低于低表达RIP1的患者(均P<0.05)。抑制RIP1基因后其相对表达量、细胞侵袭、迁移能力均下降(P<0.05)。RIP1在肝癌中存在高表达,可能参与肝癌细胞侵袭与迁移过程。
This study is aimed to investigate the expression of receptor-interacting protein kinase-1( RIP1) in hepatocellular carcinoma and its relationship with histopathology,and to explore its effect on the invasion and migration of hepatoma cells. Tumor tissues and paracancerous tissues were collected; the expression of RIP1 in the hepatocellular carcinoma cell line was screened and the cell line with the highest expression of RIP1 was collected for subsequencing experiments.The RIP1 gene was inhibited and normal control group was set. Western blotting and RT-PCR were used to detect the expression of RIP1. Tranwell was used to observe the invasive ability of the cells. Cell migration was observed in cell scratches. Of the 48 patients with liver cancer,22 were stained,and the positive rate of staining was 45.83%. Of these,15 were strongly positive,7 were moderately and weakly positive,and 6 were relapsed. The expression of RIP1 in hepatocellular carcinoma was significantly higher than that in paracancerous tissue( P<0.05). Total survival rate and relapse-free survival rate in patients with high expression of RIP1 was significantly higher than that in patients with low expression of RIP1( P<0.05). After inhibiting RIP1 gene,its relative expression,cell invasion and migration ability were all decreased( P<0.05). RIP1 is high expressed in liver cancer and it may participate in the invasion and migration of liver cancer cells.
This study is aimed to investigate the expression of receptor-interacting protein kinase-1( RIP1) in hepatocellular carcinoma and its relationship with histopathology,and to explore its effect on the invasion and migration of hepatoma cells. Tumor tissues and paracancerous tissues were collected; the expression of RIP1 in the hepatocellular carcinoma cell line was screened and the cell line with the highest expression of RIP1 was collected for subsequencing experiments.The RIP1 gene was inhibited and normal control group was set. Western blotting and RT-PCR were used to detect the expression of RIP1. Tranwell was used to observe the invasive ability of the cells. Cell migration was observed in cell scratches. Of the 48 patients with liver cancer,22 were stained,and the positive rate of staining was 45.83%. Of these,15 were strongly positive,7 were moderately and weakly positive,and 6 were relapsed. The expression of RIP1 in hepatocellular carcinoma was significantly higher than that in paracancerous tissue( P<0.05). Total survival rate and relapse-free survival rate in patients with high expression of RIP1 was significantly higher than that in patients with low expression of RIP1( P<0.05). After inhibiting RIP1 gene,its relative expression,cell invasion and migration ability were all decreased( P<0.05). RIP1 is high expressed in liver cancer and it may participate in the invasion and migration of liver cancer cells.
引文
[1]杜文杰,刘祖炳,刘华生.原发性肝癌介入治疗中经导管肝动脉栓塞与肝动脉灌注化疗栓塞的远期疗效比较[J].中华肿瘤防治杂志,2016(s2):169-71.
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[3]崔凯,耿慧武,姬强,等.受体相互作用蛋白激酶1在食管鳞状细胞癌中的表达及临床意义[J].安徽医科大学学报,2016,51(8):1175-9.
[4]王书乐,成祥,陈国柱,等.肿瘤坏死因子α通过抑制线粒体呼吸链复合体Ⅲ诱导L929-A细胞发生RIP1激酶依赖性细胞凋亡[J].军事医学,2017,41(5):346-51.
[5]LIN X,CHEN Q,CHEN H,et al.CYLD promotes TNF-α-induced cell necrosis mediated by RIP-1 in human lung cancer cells[J].Mediat Inflamm,2016,2016(4,article 400):1-11.
[6]WU P,SHI K J,AN J J,et al.The LEF1/CYLD axis and c IAPs regulate RIP1 deubiquitination and trigger apoptosis in selenitetreated colorectal cancer cells[J].Cell Death Dis,2014,5(2):e1085.
[7]章余妹,吴萍,张林杰,等.RIP1增强顺铂诱导食管癌细胞的凋亡[J].安徽医科大学学报,2015,(2):172-6.
[8]QIU X,KLAUSRN C,CHENG JC,et al.CD40 ligand induces RIP1-dependent,necroptosis-like cell death in low-grade serous but not serous borderline ovarian tumor cells[J].Cell Death Dis,2015,6(8):e1864.
[9]FU ZZ,DENG BY,LIAO YX,et al.The anti-tumor effect of shikonin on osteosarcoma by inducing RIP1 and RIP3 dependent necroptosis[J].BMC Cancer,2013,13:580-7.
[10]ZHU G,YE J,HUANG Y,et al.Receptor-interacting protein-1promotes the growth and invasion in gastric cancer[J].Int J Oncol,2016,48(6):2387.
[11]LIU X Y,LAI F,YAN X G,et al.RIP1 kinase is an oncogenic driver in melanoma[J].Cancer Res,2015,75(8):1736.
[12]张金灿,胡丹,贺礼进,等.干扰miR-21表达抑制胶质瘤U251细胞增殖迁移与侵袭[J].中南医学科学杂志,2016,44(2):143-6.
[13]王攀,谢敖文,范钦桥,等.miR-139-5p靶向转化生长因子-β1抑制肝癌细胞侵袭转移的机制[J].中华肝胆外科杂志,2016,22(1):17-23.
[14]别彩群,黄秋燕,颜英,等.转染沉默IGF1R基因的肝癌细胞株增殖、迁移及侵袭能力观察[J].山东医药,2016,56(11):1-4.
[15]LEI F X,JIN L,LIU X Y,et al.RIP1 protects melanoma cells from apoptosis induced by BRAF/MEK inhibitors[J].Cell Death Dis,2018,9(6):679.
[16]PIETKIEWICZ S,EILS R,KRAMMER P H,et al.Combinatorial treatment of CD95L and gemcitabine in pancreatic cancer cells induces apoptotic and RIP1-mediated necroptotic cell death network[J].Exp Cell Res,2015,339(1):1-9.
[2]黄乾荣,张玲.原发性肝癌治疗研究新进展[J].实用医学杂志,2016,32(14):2275-8.
[3]崔凯,耿慧武,姬强,等.受体相互作用蛋白激酶1在食管鳞状细胞癌中的表达及临床意义[J].安徽医科大学学报,2016,51(8):1175-9.
[4]王书乐,成祥,陈国柱,等.肿瘤坏死因子α通过抑制线粒体呼吸链复合体Ⅲ诱导L929-A细胞发生RIP1激酶依赖性细胞凋亡[J].军事医学,2017,41(5):346-51.
[5]LIN X,CHEN Q,CHEN H,et al.CYLD promotes TNF-α-induced cell necrosis mediated by RIP-1 in human lung cancer cells[J].Mediat Inflamm,2016,2016(4,article 400):1-11.
[6]WU P,SHI K J,AN J J,et al.The LEF1/CYLD axis and c IAPs regulate RIP1 deubiquitination and trigger apoptosis in selenitetreated colorectal cancer cells[J].Cell Death Dis,2014,5(2):e1085.
[7]章余妹,吴萍,张林杰,等.RIP1增强顺铂诱导食管癌细胞的凋亡[J].安徽医科大学学报,2015,(2):172-6.
[8]QIU X,KLAUSRN C,CHENG JC,et al.CD40 ligand induces RIP1-dependent,necroptosis-like cell death in low-grade serous but not serous borderline ovarian tumor cells[J].Cell Death Dis,2015,6(8):e1864.
[9]FU ZZ,DENG BY,LIAO YX,et al.The anti-tumor effect of shikonin on osteosarcoma by inducing RIP1 and RIP3 dependent necroptosis[J].BMC Cancer,2013,13:580-7.
[10]ZHU G,YE J,HUANG Y,et al.Receptor-interacting protein-1promotes the growth and invasion in gastric cancer[J].Int J Oncol,2016,48(6):2387.
[11]LIU X Y,LAI F,YAN X G,et al.RIP1 kinase is an oncogenic driver in melanoma[J].Cancer Res,2015,75(8):1736.
[12]张金灿,胡丹,贺礼进,等.干扰miR-21表达抑制胶质瘤U251细胞增殖迁移与侵袭[J].中南医学科学杂志,2016,44(2):143-6.
[13]王攀,谢敖文,范钦桥,等.miR-139-5p靶向转化生长因子-β1抑制肝癌细胞侵袭转移的机制[J].中华肝胆外科杂志,2016,22(1):17-23.
[14]别彩群,黄秋燕,颜英,等.转染沉默IGF1R基因的肝癌细胞株增殖、迁移及侵袭能力观察[J].山东医药,2016,56(11):1-4.
[15]LEI F X,JIN L,LIU X Y,et al.RIP1 protects melanoma cells from apoptosis induced by BRAF/MEK inhibitors[J].Cell Death Dis,2018,9(6):679.
[16]PIETKIEWICZ S,EILS R,KRAMMER P H,et al.Combinatorial treatment of CD95L and gemcitabine in pancreatic cancer cells induces apoptotic and RIP1-mediated necroptotic cell death network[J].Exp Cell Res,2015,339(1):1-9.
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