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目的:探讨Rce 1(Ras and a-factor converting enzyme 1)在膀胱癌组织及对应癌旁组织中的表达差异及其意义;探究慢病毒介导sh RNA沉默Rce1基因对人膀胱癌细胞株T24细胞凋亡的影响。方法:采用q RT-PCR和Western blot技术检测Rce1在12例膀胱癌组织及对应癌旁组织中m RNA和蛋白表达情况,采用免疫组化技术检测76例膀胱癌组织及对应癌旁组织中Rce1表达情况;结合临床病理资料,分析Rce1表达与临床病理特征的关系。设计针对Rce1基因的sh RNA慢病毒干扰载体感染T24细胞,经嘌呤霉素筛选后建立稳定感染细胞系,实时荧光定量PCR、Western blot分别检测感染后各组细胞Rce1 m RNA和蛋白表达水平;应用流式细胞仪检测T24细胞凋亡改变情况。结果:q RT-PCR和Western blot显示在12对癌及癌旁组织中Rce1差异性表达,差异有统计学意义(P=0.00);78例癌组织中Rce1阳性表达率为70.5%(55/78),癌旁组织中阳性率为19.0%(4/21),组间差异有统计学意义(P=0.000);统计学分析表明,Rce1表达量与肿瘤组织学分级、T分期和肿瘤是否远处转移存在相关关系(P<0.05);Kaplan-Meier生存分析显示,Rce1阳性表达与病人术后生存时间呈负相关关系(P=0.016)。与空白对照组和阴性对照组相比,感染sh RNA慢病毒干扰载体细胞组Rce1 m RNA和蛋白表达水平明显下调(P=0.000),细胞凋亡明显增加。结论:Rce1在膀胱癌组织中表达高于癌旁组织,Rce1在膀胱癌的发生发展中可能发挥重要作用;慢病毒介导sh RNA沉默Rce1基因能有效下调Rce1在T24细胞中表达,并诱导细胞凋亡;Rce1有望成为膀胱癌临床诊治和预后的一个新指标。
Objective: To investigate the expression and significance of Rce 1 (Ras and a-factor converting enzyme 1) in bladder cancer tissues and corresponding paracancerous tissues, and to explore the effect of Rce1 gene mediated by lentivirus-mediated sh RNA on human bladder cancer cell line T24 cells Effect of apoptosis. Methods: The mRNA and protein expressions of Rce1 in 12 cases of BTCC and corresponding paracancerous tissues were detected by q RT-PCR and Western blot. Immunohistochemistry was used to detect the expression of Rce1 in 76 cases of bladder cancer and corresponding paracancerous tissues The relationship between Rce1 expression and clinicopathological features was analyzed based on the clinicopathological data. T cells were infected by sh RNA lentiviral vector targeting Rce1 gene and stable infected cell lines were selected after puromycin selection. Rce1 m RNA and protein expression levels were detected by real-time fluorescence quantitative PCR and Western blot respectively. Flow cytometry T24 cells apoptosis changes. Results: RT-PCR and Western blot showed that Rce1 was differentially expressed in 12 pairs of cancerous tissues and paracancerous tissues (P = 0.00). The positive rate of Rce1 expression in 78 of cancerous tissues was 70.5% (55 / 78). The positive rate of adjacent tissues was 19.0% (4/21), with significant difference between groups (P = 0.000). Statistical analysis showed that the expression of Rce1 correlated with histological grade, T stage and tumor Distant metastasis (P <0.05). The Kaplan-Meier survival analysis showed that the positive expression of Rce1 was negatively correlated with the postoperative survival time (P = 0.016). Compared with the blank control group and the negative control group, the expression of Rce1 m RNA and protein in the vector cells transfected with sh RNA lentivirus was significantly decreased (P = 0.000), and the apoptosis rate was significantly increased. CONCLUSION: Rce1 is highly expressed in bladder cancer tissues and Rce1 may play an important role in the development of bladder cancer. Lentivirus-mediated silencing of Rce1 gene can effectively down-regulate the expression of Rce1 in T24 cells and induce cell growth Apoptosis; Rce1 is expected to become a new indicator of clinical diagnosis and prognosis of bladder cancer.