siRNA沉默钙蛋白酶1对人脑胶质瘤细胞LN229增殖及侵袭的影响  

Effect of siRNA silencing calpain 1 on proliferation and invasion of human glioma cell line LN229

作　　者：唐树军 胡峻华 施登超 宁威海 吴科 TANG Shu-jun;HU Jun-hua;SHI Deng-chao;NING Wei-hai;WU Ke(Department of Neurosurgery,People's Hospital of Xichang City,Sichuan Province 615000,China;Department of Pathology,Central Hospital of Panzhihua,Sichuan Province 617000,China;Department of Neurology,Sanbo Brain Hospital,Capital Medical University,Beijing 100093,China)

机构地区：[1]四川省西昌市人民医院神经外科,四川西昌615000 [2]四川省攀枝花市中心医院病理科,四川攀枝花617000 [3]首都医科大学三博脑科医院神经肿瘤科,北京海淀100093

出　　处：《解剖学研究》2023年第6期518-522,共5页Anatomy Research

基　　金：四川省卫生健康委医学科技项目(21PJ033)。

摘　　要：目的 探讨siRNA沉默钙蛋白酶1(CAPN1)对人脑胶质瘤细胞LN229增殖及侵袭的影响。方法 取人脑胶质瘤细胞LN229于37℃水浴中解冻后制备单细胞悬液,接种于细胞培养板中,融合度至90%左右,将siRNA系列、CAPN1 siRNA序列分别转染细胞,分为实验组(CAPN1 siRNA)与siRNA组,以未转染的细胞作为对照组。采用实时荧光定量PCR(qRT-PCR)测定LN229细胞中CAPN1 mRNA表达;采用MTT法测定LN229细胞增殖情况;采用Transwell小室测定LN229细胞侵袭及迁移情况;采用Western blot测定LN229细胞Bax、Bcl-2和CAPN1蛋白表达。结果 与对照组和siRNA组比较,实验组CAPN1 mRNA表达明显降低(P<0.05);与对照组和siRNA组比较,实验组LN229细胞增殖率明显降低(P<0.05);与对照组和siRNA组比较,实验组LN229细胞侵袭率明显降低(P<0.05);与对照组和siRNA组比较,实验组LN229细胞迁移数目明显降低(P<0.05);与对照组和siRNA组比较,实验组LN229细胞Bax蛋白表达灰度值明显升高,而Bcl-2和CAPN1蛋白表达灰度值明显降低(P<0.05)。而对照组和siRNA组CAPN1 mRNA表达、LN229细胞增殖率、LN229细胞侵袭率、LN229细胞迁移数目、Bax、Bcl-2和CAPN1蛋白表达灰度值比较差异无统计学意义(P>0.05)。结论 siRNA沉默CAPN1可抑制人脑胶质瘤细胞LN229增殖,抑制脑胶质瘤细胞侵袭和迁移,上调Bax表达及下调Bcl-2和CAPN1蛋白表达。Objective To investigate the effect of siRNA silencing calpain 1(CAPN1)on the proliferation and invasion of human glioma cell line ln229.Methods Human glioma cell line ln229 was thawed in a 37℃water bath to prepare a single cell suspension and inoculated into the cell culture plate.The fusion degree was about 90%。SiRNA series and CAPN1 siRNA sequence were transfected into the cells respectively.The cells were divid⁃ed into experimental group(CAPN1 siRNA)and siRNA group,and the non transfected cells were used as the con⁃trol group.The expression of CAPN1 mRNA in LN229 cells was measured by real⁃time fluorescence quantitative PCR(QRT PCR);the proliferation of LN229 cells was detected by MTT assay;the invasion and migration of LN229 cells were measured by Transwell chamber;the expressions of Bax,Bcl⁃2 and CAPN1 proteins in LN229 cells were measured by Western blot.Results Compared with the control group and siRNA group,the expression of CAPN1 mRNA in the experimental group decreased significantly(P<0.05);Compared with the control group and siRNA group,the proliferation rate of LN229 cells in the experimental group decreased significantly(P<0.05);Compared with the control group and siRNA group,the invasion rate of LN229 cells in the experimental group was significantly lower(P<0.05);Compared with the control group and siRNA group,the migration number of LN229 cells in the experimental group decreased significantly(P<0.05);Compared with the control group and siRNA group,the gray value of Bax protein expression in LN229 cells in the experimental group was significantly higher,while the gray value of Bcl⁃2 and CAPN1 protein expression was significantly lower(P<0.05).There was no signifi⁃cant difference in CAPN1 mRNA expression,LN229 cell proliferation rate,LN229 cell invasion rate,LN229 cell migration number,Bax,Bcl⁃2 and CAPN1 protein expression gray value between the control group and siRNA group(P>0.05).Conclusion siRNA silencing CAPN1 can inhibit the proliferation of human glioma cell line ln229,in

关 键 词：钙蛋白酶1 人脑胶质瘤 SIRNA 增殖 侵袭 LN229细胞 

分 类 号：R739.41[医药卫生—肿瘤]