[关键词]
[摘要]
目的:探讨富脯氨酸蛋白11(PRR11)在食管癌组织中的表达及其在体外对人食管癌TE-2 细胞侵袭和迁移能力的影响。方法:选取2016 年10 月至2018 年10 月郑州大学第二附属医院胸外科经病理确诊为食管癌患者80 例,收集其手术切除的食管癌组织及相应的癌旁组织标本。用qPCR检测食管癌组织或细胞系中PRR11 mRNA的表达水平,Log-rank Test 法分析食管癌组织中PRR11 mRNA的表达与患者一般资料、组织学分级、淋巴结转移、浸润深度及TNM分期的关系,Kaplan-Meier 曲线分析法分析PRR11 mRNA与食管癌患者预后的关系。以慢病毒shRNA表达载体感染食管癌TE-2 细胞,构建敲低PRR11 的细胞系及相应的对照细胞系作为本研究的shPRR11#1、shPRR11#2 及对照组,qPCR及WB实验分别验证细胞株中PRR11 mRNA及蛋白的表达水平,MTT实验检测感染后细胞的增殖能力,Transwell 实验检测其侵袭和迁移能力。结果:PRR11 mRNA在食管癌组织中的表达高于癌旁组织(P<0.05),PRR11 mRNA的过表达与食管癌的组织学分级、淋巴结转移、浸润深度及TNM分期均显著相关(均P<0.05),PRR11 高表达与食管癌患者预后不良显著相关(P<0.05);shPRR11#1、shPRR11#2 组细胞中PRR11 mRNA及蛋白表达显著低于对照组细胞(P<0.05 或P<0.01)。shPRR11#1、shPRR11#2 组细胞的增殖能力低于对照组(P<0.05 或P<0.01),Transwell侵袭及迁移实验结果显示,shPRR11#1、shPRR11#2 组平均每个视野内细胞数均显著低于对照组(P<0.01)。结论:PRR11 高表达于食管癌组织中,与食管癌的发生、进展及患者预后密切相关;体外实验也证明了敲低PRR11 表达可以抑制食管癌的增殖、侵袭和迁移能力,是潜在的食管癌靶标。
[Key word]
[Abstract]
Objective: To investigate the expressionof proline-rich protein 11 (PRR11) in esophageal carcinoma (EC) tissues and to study it’s effect on the proliferation and metastasis of human EC TE-2 cells in vitro. Methods: Eighty patients were pathologically diagnosed with EC the Department of Thoracic Surgery of the Second Affiliated Hospital of Zhengzhou University from October 2016 to October 2018, and their surgically resected cancer tissues and corresponding para-cancerous tissues were collected for this study. qPCR was used to detect the expression of PRR11 mRNA in tissues or cells. Log-rank Test was used to analyzethe relationship between the expression of PRR11 in EC tissues and general data, histological type, lymphatic metastasis, depth of invasion and TNM stageof the EC patients. Kaplan-Meierplot was used to analyze the association between PRR11 mRNA and patients’prognosis. TE-2 cells were transfected with lentivirus shRNA to construct cell line with PRR11 knockout and corresponding control cell lines, as shPRR11#1,shPRR11#2 and control group. qPCR and WB assays were used to verify the mRNA and protein expressions of PRR11 in cell lines respectively.MTT was used to examine the proliferation of transfected cells, and Transwell experiments were used to detect cell invasion and migration. Results: The expression of PRR11 mRNA in EC was higher than that in para-cancer tissues (P<0.05). There was sig‐nificant correlation between PRR11 over-expression and histological type, lymphatic metastasis, depth of invasion and TNM stage(all P <0.05), and high PRR11 expression was significantly related with the poor prognosis of EC patients (P<0.05). The mRNA and protein expressions of PRR11 in cells of shPRR11#1 and shPRR11#2 groups were significantly lower than those in control group (all P<0.05).MTT assay showed that the proliferation of cells in shPRR11#1 and shPRR11#2 groups was significantly lower than that in the control group (P<0.05 or P<0.01). The results of Transwell invasion and migration assays showed that the average number of cells with in each field of viewin shPRR11#1 and shPRR11#2 groups was significantly lower than that in the control group (P<0.01). Conclusion: PRR11 is over-expressed in EC tissues and PRR11 over-expression is closely related to the occurrence, progression and prognosis of esophageal cancer. In vitro experiments have also demonstrated that knockdown of PRR11 can inhibit the proliferation, invasion and migration of EC. PRR11 can be used as a potential molecule marker and drug targets for EC.
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[基金项目]
河南省高等学校青年骨干教师培养计划资助项目(No.2016GGJS-261);河南省科技攻关资助项目(No.192102310103)