CLIC1对氯化钴诱导的胃癌细胞增殖侵袭及MAPK/ERK通路的影响
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摘要
目的:探讨CLIC1对氯化钴诱导的胃癌细胞增殖、侵袭及MAPK/ERK通路的影响。方法:体外培养胃癌细胞SGC-7901后分为4组,正常组(control组)、氯化钴处理组(150um CoCl2处理);阴性转染组(si-NC组,CoCl2处理后继续培养48h,转染NC siRNA); CLIC1干扰组(si-CLIC1组,CoCl2处理后继续培养48h,转染CLIC1siRNA)。收集细胞上清,MTT法检测SGC-7901细胞活力;平板细胞克隆形成实验检测菌落形成;划痕实验检测细胞转移能力; Transwell小室检测细胞迁移、侵袭能力;蛋白免疫印迹(WB)检测MAPK/ERK通路有关蛋白的表达。结果:与control组相比,CoCl两组、si-NC组细胞抑制率、转移抑制率、E-cadherin蛋白表达均升高,菌落数量、迁移、侵袭数量均降低,p-ERK1/2、MMP-9、N-cadherin蛋白表达降低(P <0.05),抑制CLIC1表达后细胞抑制率、转移抑制率、E-cadherin蛋白表达进一步升高,菌落数量、迁移、侵袭数量以及p-ERK1/2、MMP-9、N-cadherin蛋白表达进一步降低(P<0.05)。结论:CoCl2处理后可抑制胃癌细胞的增殖、侵袭,而抑制CLIC1表达后能够协同CoCl2进一步抑制胃癌细胞的增殖、侵袭,这可能与抑制MAPK/ERK通路有关。
Objective: To study the effects of CLIC1 on the proliferation,invasion and MAPK/ERK pathway of gastric cancer cells induced by cobalt chloride,so as to provide certain ideas for the treatment of diseases. Methods: After cultured gastric cancer cells SGC-7901 in vitro,they were divided into 4 groups,the normal group(group control) and the cobalt chloride treatment group(150 um CoCl2 treatment); in the negative transfection group(group si-NC,CoCl2 was treated with 48 h and transfected with NC siRNA);CLIC1 interference group(group si-CLIC1,CoCl2 after treatment continued to cultivate 48 h,transfected CLIC1 siRNA). The cell supernatants were collected,the viability of SGC-7901 cells was detected by MTT;colony formation assay was used to detect colony formation; the scratch test was used to detect the cell transfer ability; transwell cell was used to detect cell migration and invasion; Western blot(WB) was used to detect the expression of MAPK/ERK pathway related proteins. Results: Compared with control group,the inhibition rate,metastasis inhibition rate and E-cadherin protein expression in CoCl2 group and si-NC group increased,the number of colonies,migration and invasion decreased,while the expression of p-ERK1/2,MMP-9 and N-cadherin protein decreased(P <0.05). After inhibiting CLIC1 expression,the inhibition rate,metastasis inhibition rate and E-cadherin protein expression increased further,and the number,migration and invasion of colonies increased. The number and expression of p-ERK1/2,MMP-9 and N-cadherin protein were further decreased(P <0.05). Conclusion: CoCl2 treatment can inhibit the proliferation and invasion of gastric cancer cells,and the inhibition of CLIC1 expression can further inhibit the proliferation and invasion of gastric cancer cells in coordination with CoCl2,which may be related to the inhibition of MAPK/ERK pathway.
Objective: To study the effects of CLIC1 on the proliferation,invasion and MAPK/ERK pathway of gastric cancer cells induced by cobalt chloride,so as to provide certain ideas for the treatment of diseases. Methods: After cultured gastric cancer cells SGC-7901 in vitro,they were divided into 4 groups,the normal group(group control) and the cobalt chloride treatment group(150 um CoCl2 treatment); in the negative transfection group(group si-NC,CoCl2 was treated with 48 h and transfected with NC siRNA);CLIC1 interference group(group si-CLIC1,CoCl2 after treatment continued to cultivate 48 h,transfected CLIC1 siRNA). The cell supernatants were collected,the viability of SGC-7901 cells was detected by MTT;colony formation assay was used to detect colony formation; the scratch test was used to detect the cell transfer ability; transwell cell was used to detect cell migration and invasion; Western blot(WB) was used to detect the expression of MAPK/ERK pathway related proteins. Results: Compared with control group,the inhibition rate,metastasis inhibition rate and E-cadherin protein expression in CoCl2 group and si-NC group increased,the number of colonies,migration and invasion decreased,while the expression of p-ERK1/2,MMP-9 and N-cadherin protein decreased(P <0.05). After inhibiting CLIC1 expression,the inhibition rate,metastasis inhibition rate and E-cadherin protein expression increased further,and the number,migration and invasion of colonies increased. The number and expression of p-ERK1/2,MMP-9 and N-cadherin protein were further decreased(P <0.05). Conclusion: CoCl2 treatment can inhibit the proliferation and invasion of gastric cancer cells,and the inhibition of CLIC1 expression can further inhibit the proliferation and invasion of gastric cancer cells in coordination with CoCl2,which may be related to the inhibition of MAPK/ERK pathway.
引文
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[9]刘晓玲,包韩乌云,赵华路,等.在Co Cl2模拟低氧条件下HIF-1α直接调控过氧化物酶体增殖物激活受体γ2(PPARγ2)的表达[J].基础医学与临床,2015,35(5):585~589.
[10]张宾,李畅,贾雪峰,等.子宫内膜样腺癌中CLIC1蛋白的表达及临床意义[J].临床肿瘤学杂志,2014,19(1):49~51.
[11] Ding Q,Li M,Wu X,et al. CLIC1 overexpression is associated with poor prognosis in gallbladder cancer[J].Tumour Biology the Journal of the International Society for Oncodevelopmental Biology&Medicine,2015,36(1):193~198.
[12] Li B P,Mao Y T,Wang Z,et al. CLIC1 Promotes the Progression of Gastric Cancer by Regulating the MAPK/AKT Pathways.[J]. Cellular Physiology&Biochemistry,2018,46(3):907~924.
[13]王妍,刘云鹏,曲秀娟,等.RANKL通过激活Akt及ERK介导胃癌细胞迁移[J].中国医科大学学报,2013,42(7):591~594.
[14]马营营,黄煜,颜莉莉,等.ERK信号转导通路在CXCL12促进子宫内膜癌细胞增殖和侵袭中的作用[J].中国肿瘤生物治疗杂志,2016,23(2):250~254.
[2]伍友兴,桂若虎,郑新平,等.水飞蓟宾抑制缺氧状态下胃癌细胞m TOR磷酸化及HIF-1α表达[J].中国肿瘤生物治疗杂志,2016,23(1):52~56.
[3] Ding Q,Li M,Wu X,et al. CLIC1 overexpression is associated with poor prognosis in gallbladder cancer[J]. Tumour Biology the Journal of the International Society for Oncodevelopmental Biology&Medicine,2015,36(1):193~198.
[4] Zhao W,Mingshu L U,Zhang Q. Chloride intracellular channel 1 regulates migration and invasion in gastric cancer by triggering the ROS-mediated p38 MAPK signaling pathway:[J]. mo Lecular Medicine Reports,2016,12(6):8041~8047.
[5] Lu J,Dong Q,Zhang B,et al. Chloride intracellular channel1(CLIC1)is activated and functions as an oncogene in pancreatic cancer[J]. Medical Oncology,2015,32(6):616~622.
[6]郭洋,吴密璐,赵君慧,等.肺癌低氧微环境与化疗耐药相关性研究进展[J].中国肺癌杂志,2014,17(3):265~268.
[7]王育蓉,李艳艳,卢仁隆.Sp1在低氧肝癌细胞血管内皮生长因子转录调控中的作用[J].军事医学,2017,41(7):572~575.
[8]徐峥,陈武桂,孙靖,等.低氧抑制乳腺癌细胞中信号素3A表达并调控成骨前体细胞分化[J].肿瘤,2016,36(12):1298~1306.
[9]刘晓玲,包韩乌云,赵华路,等.在Co Cl2模拟低氧条件下HIF-1α直接调控过氧化物酶体增殖物激活受体γ2(PPARγ2)的表达[J].基础医学与临床,2015,35(5):585~589.
[10]张宾,李畅,贾雪峰,等.子宫内膜样腺癌中CLIC1蛋白的表达及临床意义[J].临床肿瘤学杂志,2014,19(1):49~51.
[11] Ding Q,Li M,Wu X,et al. CLIC1 overexpression is associated with poor prognosis in gallbladder cancer[J].Tumour Biology the Journal of the International Society for Oncodevelopmental Biology&Medicine,2015,36(1):193~198.
[12] Li B P,Mao Y T,Wang Z,et al. CLIC1 Promotes the Progression of Gastric Cancer by Regulating the MAPK/AKT Pathways.[J]. Cellular Physiology&Biochemistry,2018,46(3):907~924.
[13]王妍,刘云鹏,曲秀娟,等.RANKL通过激活Akt及ERK介导胃癌细胞迁移[J].中国医科大学学报,2013,42(7):591~594.
[14]马营营,黄煜,颜莉莉,等.ERK信号转导通路在CXCL12促进子宫内膜癌细胞增殖和侵袭中的作用[J].中国肿瘤生物治疗杂志,2016,23(2):250~254.