摘要
目的探讨蛋白质精氨酸甲基转移酶1(PRMT1)结合羟甲基戊二酸单酰辅酶A还原酶(HMGCR)后活性变化及其对食管鳞癌细胞增殖能力的影响。方法以食管鳞癌ECA109细胞为模型,通过免疫共沉淀法观察PRMT1与HMGCR在细胞中的相互结合作用。将ECA109细胞随机分为感染组和对照组,分别以表达特异性针对PRMT1的RNA干扰分子的慢病毒质粒转染48 h,采用Western blot法分别检测PRMT1、HMGCR蛋白及甲基化HMGCR蛋白,分光光度法检测HMGCR酶活性,CCK-8法检测细胞增殖活性。结果细胞裂解后的蛋白共沉淀复合物分别用抗PRMT1、HMGCR抗体捕获后,可检测到HMGCR、PRMT1蛋白条带。感染组和对照组ECA109细胞中PRMT1蛋白相对表达量分别为0.057±0.011和0.239±0.041(P<0.01),HMGCR蛋白相对表达量分别为0.175±0.031和0.184±-0.037(P>0.05)。感染组和对照组ECA109细胞甲基化HMGCR蛋白相对表达量分别为0.028±0.006、0.107±0.016,HMGCR相对活性分别为0.104±0.009、0.277±0.013,P均<0.01。感染组ECA109细胞培养24 h后,细胞增殖的OD_(450)值已低于对照组(P<0.05);培养48 h及72 h后,两组OD_(450)值差距进一步扩大(P均<0.01)。结论 PRMT1可通过结合HMGCR蛋白促进其甲基化途径增强HMGCR蛋白活性,进而促进食管鳞癌细胞的体外增殖能力。
Objective To investigate the effect of protein arginie methyltransferase 1 （ PRMT1 ） interacting with HMG-CoA reductase （HMGCR） and regulating its activity on the proliferation of esophageal squamous cell carcinoma （ES- CC） cells. Methods Human esophageal cancer ECA109 cells were chosen as the cell models. The interaction between PRMT1 and HMGCR was detected by co-immunoprecipitation. ECA109 cells were randomly divided into two groups： the infection group and the control group. ECA109 cells were infected with lentivirus expressing small hairpin RNA of HMGCR for 48 h. Then, the relative expression level of PRMT1, total and methylated HMGCR protein was detected by Western blotting. The relative activity of HMGCR was detected by ultraviolet spectrophotometry. Cell proliferation was detected by CCK-8 assay. Results In the infection group and control group, the protein expression of PRMT1 in ECA109 cells of was 0. 057 ＋0. 011 and 0. 239 ＋0. 041, respectively （P 〈0.01 ） , and the protein expression of HMGCR was 0. 175 ±0. 031 and 0. 184 ±0. 037 （P 〉 0.05 ）. In ECA109 cells of the infection group and control group, the protein expression of meth- ylated HMGCR was 0.028 ± 0.006 and 0. 107± 0. 016, and the relative activity was 0. 104 ± 0.009 and 0. 277 ±0. 013, respectively, （all P 〈 0.01 ）. The OD45 of ECA109 cells after 24-hour culture in the infection group was significantly lower than that of control group （ P 〈 0.05 ） , and after 48 h and 72 h, the difference between these two groups further expanded （ all P 〈 0.01 ）. Conclusion PRMT1 increases the proliferative capacity of ESCC cells by interacting with HMGCR and regulating its activity.;： esophageal squamous carcinoma; protein arginie methyltransferase 1; HMG-CoA reductase; cell proliferation
出处
《山东医药》
CAS
北大核心
2016年第38期19-22,共4页
Shandong Medical Journal
基金
河南省科技攻关计划项目(122102310618)
