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目的:探讨nm23-H1基因稳定表达对人宫颈细胞增殖能力的影响,及其表达与临床特点的关系。方法:用免疫组织化学法检测nm23-H1基因在宫颈癌组织的表达。将真核表达载体pcDNA3.1-nm23-H1转染入宫颈癌细胞系HeLa、SiHa,G418稳定筛选,用Western印迹法、免疫组化法鉴定转染前后细胞中nm23-H1基因的蛋白表达,噻唑蓝(MTT)法检测转染前后细胞体外增殖能力的变化。结果:在宫颈鳞癌组织中,nm23-H1基因阳性表达率随FIGO临床分期的升高有下降趋势,Ⅲ期nm23-H1阳性表达率明显低于Ⅱ期,Ⅱ期阳性表达率明显低于I期(P<0.01);分化程度高的组织阳性表达率显著高于分化程度低的组织(P<0.01);有淋巴结转移者其原发灶阳性表达率明显低于无淋巴结转移者(P<0.01)。宫颈腺癌组织中nm23-H1基因阳性表达率与FIGO临床分期,分化程度,淋巴结转移均无明显的相关性(P>0.05)。在pcDNA3.1-nm23-H1转染组中,Si-Ha、HeLa细胞nm23-H1蛋白表达水平明显增加,未转染组和空载体对照组未见nm23-H1高表达。MTT法所绘生长曲线显示,SiHa-nm23(转染组)细胞与SiHa-3.1(空载体组)、SiHa(空白组)细胞比较,生长速度明显抑制,差异均有统计学意义(P<0.01),而HeLa(空白组)、HeLa-nm23(转染组)、HeLa-3.1(空载体组)三组细胞生长速度均无明显变化(P>0.05)。结论:nm23-H1参与了宫颈鳞癌的发生、发展过程,是判断宫颈鳞癌淋巴结转移和预后的重要标志物,而nm23-H1基因对不同宫颈癌细胞体外增殖能力的影响有明显差异。
Objective: To investigate the effect of stable expression of nm23-H1 gene on human cervical cell proliferation and its relationship with clinical features. Methods: The expression of nm23-H1 gene in cervical cancer was detected by immunohistochemistry. The eukaryotic expression vector pcDNA3.1-nm23-H1 transfected into cervical cancer cell lines HeLa, SiHa, G418 stable screening, Western blotting, immunohistochemical staining of nm23-H1 gene expression in cells before and after transfection, The MTT assay was used to detect the proliferation of cells before and after transfection. Results: In cervical squamous cell carcinoma, the positive rate of nm23-H1 gene decreased with the increase of FIGO clinical stage, the positive rate of nm23-H1 in stage Ⅲ was significantly lower than that of stage Ⅱ, the positive rate of nm23-H1 in stage Ⅱ was significantly lower than that of I (P <0.01). The positive expression rate of the tissue with high degree of differentiation was significantly higher than that with the low degree of differentiation (P <0.01). The positive rate of the primary tumor with lymph node metastasis was significantly lower than that without lymph node metastasis (P < 0.01). The positive rate of nm23-H1 gene in cervical adenocarcinoma was not correlated with clinical stage, differentiation, lymph node metastasis (FIGO) (P> 0.05). In the pcDNA3.1-nm23-H1 transfection group, the expression of nm23-H1 protein in Si-Ha and HeLa cells was significantly increased, while no expression of nm23-H1 was found in untransfected and empty vector control groups. The growth curve of MTT assay showed that the growth rate of SiHa-nm23 cells was significantly inhibited compared with SiHa-3.1 (empty vector group) and SiHa (blank group) cells (P <0.01 ), While there was no significant change in the cell growth rate in HeLa (blank group), HeLa-nm23 (transfected group) and HeLa-3.1 (empty vector group) (P> 0.05). Conclusion: nm23-H1 is involved in the occurrence and development of cervical squamous cell carcinoma, which is an important marker for determining lymph node metastasis and prognosis of cervical squamous cell carcinoma. The effect of nm23-H1 gene on the proliferation of cervical cancer cells in vitro is obviously different.