VDAC1在子宫颈鳞癌中的生物学特性及其与HPV16感染的关系

Effect of VDAC1 on biological characteristics of cervical squamous cell carcinoma and its relationship with HPV16 infection

目的探讨应用IHC和RT-PCR方法检测电压依赖型阴离子孔道蛋白(Voltage-dependent anion channel1;VDAC1)与mRNA在人子宫颈鳞癌(CSCC)中的表达与临床病理参数相关性,及其与人乳头状瘤病毒(human papillomavirus 16,HPV16)感染的关系。方法用实时定量PCR(RT-PCR)法检测新鲜组织(子宫颈鳞癌、高级别上皮内瘤变和正常组织)中VDAC1的mRNA表达;用免疫组织化学(IHC)法检测石蜡组织标本中VDAC1蛋白的表达,并分析VDAC1的表达水平与子宫颈鳞癌患者临床病理特征的相关性;用免疫印迹法(WB)检测SiHa、C33A和Ect1细胞系中VDAC1蛋白的表达。结果RT-PCR结果显示子宫颈鳞癌、高级别上皮内瘤变和正常组织中VDAC1的相对表达量分别为5.06±7.65、2.46±3.22、1.14±1.25;IHC结果显示子宫颈鳞癌、高级别上皮内瘤变和正常石蜡包埋组织中VDAC1的阳性率分别为75.0%、41.03%、0%,统计学分析VDAC1的表达与患者AJCC分期(P=0.002)、浸润深度(P=0.021)、淋巴结转移(P=0.0001)、分化程度(P=0.0144)、血管侵犯(P=0.001)、神经侵犯(P=0.0005)等临床病理参数显著相关;VDAC1蛋白在Si Ha、C33 A、Ect1细胞系中的相对表达量分别为0.92±0.50、0.50±0.36、0.23±0.05,且SiHa和Ect1(P=0.033)细胞间VDAC1的表达有统计学差异。结论VDAC1基因的高表达可能促进子宫颈鳞癌的发生、发展,并与部分临床病理参数相关;HPV16阳性的肿瘤细胞中VDAC1的表达相对较高,提示与VDAC1基因具有一定的相关性。

Objective To investigate the correlation of the expression of voltage dependent anion channel 1(VDAC1)protein and mRNA in human cervical squamous cell carcinoma(CSCC)by IHC and RT-PCR and its clinicopathological parameters with HPV16 infection.Method VDAC1 mRNA expression in fresh tissues(CSCC,CIN and normal tissues)was detected by real-time quantitative PCR(RT-PCR);VDAC1 protein expression in paraffin embedded tissues was detected by immunohistochemistry(IHC)and the correlation between the expression level of VDAC1 and the clinicopathological characteristics was analyzed;VDAC1 protein expression in SiHa,C33A and Ect1 cell lines was detected by Western blot(WB).Results The results of RT-PCR showed that the relative expression of VDAC1 was 5.06±7.65 in CSCC,2.46±3.22 in CIN and 1.14±1.25 in normal tissues;the results of IHC showed that the positive rates of VDAC1 were 75.0%in CSCC,41.03%in CIN and 0%in normal paraffin embedded tissues;the expression of VDAC1 was significantly associated with patients’AJCC stage(P=0.002),infiltration(P=0.021),lymph node metastasis(P=0.0001),differentiation(P=0.0144),vascular invasion(P=0.001),and nerve invasion(P=0.0005);the relative expression of VDAC1 protein in SiHa,C33A and Ect1 cell lines was 0.92±0.50,0.50±0.36 and 0.23±0.05,respectively.The expression of VDAC1 was statistically different between Si Ha and Ect1(P=0.033).Conclusion The high expression of VDAC1 may promote the occurrence and development of CSCC,and the high expression of VDAC1 gene is related to some clinicopathological parameters;the expression of VDAC1 in HPV16 positive tumor cells is relatively high,suggesting that there is a certain correlation between VDAC1 gene and HPV16 positive tumor cells.