目的 探究核糖体结合糖蛋白2(RPN2)在膀胱癌中的表达情况及其对膀胱癌细胞自噬的影响.方法 通过 Oncomine、UALCAN、GEPIA 2.0 和 Human Protein Atlas(HPA)数据库分析RPN2在膀胱癌组织与正常膀胱组织中的表达差异,探讨RPN2表达与膀胱癌患者的临床病理参数的关系及其对患者生存期的影响;通过String数据库进行RPN2的KEGG和GO功能富集分析,通过GEPIA数据库分析RPN2与自噬分子LAMP2的相关性;采用实时荧光定量PCR测定RPN2在不同膀胱癌细胞水平的表达;通过T24细胞水平干扰RPN2表达后,使用透射电镜观察膀胱癌T24细胞自噬超微结构变化,采用CCK-8评价细胞增殖能力,采用免疫印迹检测自噬相关蛋白LC3-Ⅱ的表达变化.结果 Oncomine、HPA、UALCAN数据库的数据分析结果和细胞水平的实验结果均证实RPN2 mRNA和蛋白表达水平在膀胱癌组织中异常升高;进一步运用UALCAN分析发现RPN2的表达与膀胱癌的淋巴结转移、肿瘤分期呈正相关,GEPIA 2.0数据库分析结果显示RPN2高表达组患者的总生存期和无病生存期均明显短于低表达组(均P＜0.05);String数据库分析发现RPN2在KEGG信号通路和GO功能富集分析中与N-糖基化作用以及内质网功能密切相关,且与LAMP2的表达呈正相关(P＜0.001).细胞水平实验结果表明,干扰RPN2的表达能够抑制T24细胞的增殖和自噬,下调自噬下游分子LC3-Ⅱ的表达(P＜0.05).结论 RPN2在膀胱癌组织中异常高表达,与膀胱癌患者总生存周期和无病生存期呈负相关,细胞水平调节膀胱癌细胞自噬,提示RPN2可能参与膀胱癌发生发展的重要诊断和治疗靶点.
The Expression of RPN2 in bladder cancer and its effect on autophagy of bladder cancer T24 cells
Objective To investigate the expression of ribophorin Ⅱ(RPN2)in bladder canc-er and its effect on autophagy in bladder cancer cells.Methods Oncomine,UALCAN,GEPIA 2.0 and Human Protein Atlas(HPA)database were used to analyze the differential expression of RPN2 in bladder cancer and normal bladder tissues,the relationship between RPN2 expression and clinicopath-ological parameters of bladder cancer and the effect of RPN2 expression on survival of bladder cancer patients.The KEGG pathway and GO function enrichment analysis were conducted to evaluate RPN2 gene by String database,the association between RPN2 and LAMP2 was performed using GEPIA.Fur-thermore,quantitative PCR was used to determine the mRNA level of RPN2 in different bladder cancer cells.After interfering RPN2 expression in T24 cells,the autophagy ultrastructure of bladder cancer T24 cells was observed by transmission electron microscopy,cell proliferation ability was evaluated by CCK-8,and the expression of autophagy associated protein LC3-Ⅱ was detected by Western blot.Re-sults The data of Oncomine,HPA,UALCAN database and cell level experiment showed that the mRNA and protein expression levels of RPN2 in bladder cancer were obviously higher than that in nor-mal bladder.UALCAN analysis displayed that RPN2 expression was positively correlated with lymph node metastasis and tumor stage of bladder cancer.GEPIA 2.0 data showed that the overall survival and disease free survival of bladder patients in the high RPN2 expression group was significantly shorter than that in the low expression group(all P＜0.05).String database analysis showed that RPN2 was closely related to N-glycoylation and endoplasmic reticulum function in KEGG signaling pathway and GO functional enrichment analysis,and was positively correlated with LAMP2 expression(P＜0.001).The results of cell level experiments showed that interfering RPN2 expression could inhibit the proliferation and autophagy of T24 cells,and down-regulate the expression of LC3-Ⅱ,a downstream molecule of autophagy(P＜0.05).Conclusions Data according to cellular and biological informa-tion database shows that RPN2 is highly expressed in bladder cancer tissues,and its expression level is negatively correlated with the overall survival and disease free survival of bladder cancer patients,and can mediate the autophagy function of bladder cancer,which demonstrate that RPN2 may be a key tar-get for diagnosis and treatment in the development of bladder cancer.
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