首页> 中文期刊> 《中国病理生理杂志》 >蛋白基因产物9.5与宫颈癌临床病理表现的关系及其作用机制的实验研究

蛋白基因产物9.5与宫颈癌临床病理表现的关系及其作用机制的实验研究

             

摘要

AIM: To detect the protein expression of protein gene product (PGP9.5) in cervical carcinoma samples, and to explore its relationship with clinicopathological features and prognosis of cervical carcinoma patients.The potential value of PGP9.5-siRNA in the treatment of cervical cancer was preliminarily investigated in vitro.METHODS:The clinical data of cervical cancer patients ( n=180 ) who received surgical treatment in Department of Gynecology of Tianjin Fifth Central Hospital and Tianjin Central Hospital of Gynecology and Obstetrics from January 2008 to June 2015 were retrospectively analyzed.Immunohistochemical staining for PGP9.5 in all pathological specimens from cervical cancer biopsy was performed.The patients were divided into high expression of PGP9.5 group and low expression of PGP9.5 group.The relationship between PGP9.5 expression and clinicopathological parameters, such as age, HPV infection, path-ological grade, tumor diameter, lymph node metastasis, depth of invasion and clinical stage, were analyzed.The overall survival was analyzed by Kaplan-Meier method and the log-rank test.The PGP9.5-siRNA, NC-siRNA, PGP9.5 eukaryotic expression plasmid and empty vector were transfected into the SiHa cells, and the effects of PGP9.5 expression on the abili- ties of colony formation and cell invasion were determined by Western blot, colony formation assay and Transwell experi-ment.RESULTS:The relationships between the expression level of PGP9.5 and patients’ pathological characteristics in-cluding grade, tumor size, lymph node metastasis, depth of invasion, vascular involvement and clinical stage were statisti-cally significant ( P<0.05) .The overall survival rates of 3 and 5 years in PGP9.5 high expression group were significantly lower than those in PGP9.5 low expression group ( P<0.05 ) .Compared with control group, the protein expression of PGP9.5, the number of colony formation and the number of invasive cells in si-PGP9.5 group were significantly decreased. The protein expression of PGP9.5, the number of colony formation and the number of invasive cells in PGP9.5 group were significantly increased.CONCLUSION:Over-expression of PGP9.5 protein indicates poor prognosis of cervical cancer, which may be a good predictor for the prognosis of cervical cancer patients.Inhibition of PGP9.5 expression may be an ef-fective way of gene therapy for cervical cancer.%目的:研究蛋白基因产物9.5(PGP9.5)在宫颈癌病理标本中的表达与宫颈癌临床病理特征及预后的关系,并初步探讨利用PGP9.5-siRNA靶向治疗宫颈癌的潜在价值。方法:回顾性分析2008年1月~2015年6月在天津市第五中心医院妇科及天津市中心妇产科医院接受手术治疗并经病理确诊的180例宫颈癌患者临床资料。所有患者宫颈癌病理标本制作病理切片并进行SP法免疫组化染色。以PGP9.5染色后评分为依据将患者分为PGP9.5高表达组和PGP9.5低表达组。观察2组患者年龄、HPV感染、病理分级、肿瘤直径、淋巴结转移、浸润深度、累及脉管和临床分期等临床病理学参数的关系。采用Kaplan-Meier法和log-rank检验进行生存分析。采用PGP9.5-siRNA、NC-siRNA、PGP9.5真核表达质粒和对照空质粒按照脂质体载体试剂说明书转染SiHa细胞,设立si-PGP9.5组、NC组、PGP9.5组和vector组。同时设立SiHa细胞空白对照( control)组。分别采用Western blot 实验、平板克隆形成实验和Transwell实验分析5组细胞PGP9.5蛋白的表达水平、克隆形成能力和侵袭能力。结果:2组患者病理分级、肿瘤直径、淋巴结转移、浸润深度、累及脉管和临床分期等临床病理参数与PGP9.5表达水平有关。 PGP9.5高表达组患者3、5年总体生存率明显低于PGP9.5低表达组患者。与control组相比,si-PGP9.5组Si-Ha细胞中PGP9.5蛋白的表达量明显降低,克隆形成数量明显减少,过膜细胞数量明显减少;PGP9.5组SiHa细胞中PGP9.5蛋白的表达量明显升高,克隆形成数量明显增多,过膜细胞数量明显增加。结论: PGP9.5蛋白高表达预示宫颈癌患者预后不良,可能成为预测宫颈癌患者预后的良好指标,对其表达进行抑制可能实现对宫颈癌的有效基因治疗。

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