高危型人乳头瘤病毒感染的宫颈脱落细胞中HPVL1蛋白表达及hTERC基因的扩增与宫颈病变的相关性研究

HPVL1 protein, application of h TERC in HR-HPV positive cervical exfoliated cytology

目的探讨高危型人乳头瘤病毒感染的宫颈脱落细胞中人乳头瘤病毒(HPV)L1蛋白表达、人类染色体端粒酶RNA(hTERC)基因的表达与宫颈病变的相关性。方法选择宫颈高危型HPV感染阳性的240例妇女的宫颈脱落细胞标本作为研究对象,240例病例按照TBS(the Bethesda system)2001年宫颈细胞学诊断系统分为以下五组:(1)细胞学检查正常及慢性炎即正常宫颈组织(NILM)组(66例);(2)宫颈不典型鳞状细胞(ASCUS)组(27例);(3)宫颈低度病变(LISL)组(13例);(4)宫颈高度病变(HISL)组(86例);(5)宫颈癌(SCC)组(48例);根据阴道镜下活检送病理组织学检查的结果分为以下5组:(1)正常宫颈或慢性宫颈炎组即NILM组(51例);(2)宫颈上皮内瘤变1级(CIN1)组(42例);(3)宫颈上皮内瘤变2级(CIN2)组(28例);(4)宫颈上皮内瘤变3级(CIN3)组(73例);(5)SCC组(46例)。通过免疫细胞化学法检测宫颈脱落细胞中HPVL1蛋白的表达,通过荧光原位杂交(FISH)技术检测hTERC基因的表达,并对HPVL1蛋白及hTERC基因在不同级别病变的宫颈脱落细胞中的表达情况进行统计学分析。结果 240例高危型HPV感染妇女中150例hTERC基因扩增阳性,hTERC基因扩增阳性率为62.5%。NILM组、CIN1组、CIN2、CIN3和SCC组hTERC基因的扩增阳性率分别为7.8%、16.7%、75.0%、98.6%和100%,五组间hTERC基因扩增阳性率之间的差异有统计学意义(χ2=131.9,P<0.001)。此五组hTERC基因扩增阳性率的大小顺序为:SCC组≈CIN3组>CIN2组>CIN1组≈NILM组。通过Spearman等级相关分析,发现随着宫颈病变恶性程度的增加,各组中hTERC基因的扩增阳性率有增加的趋势(相关系数rs=0.71,P<0.001)。240例高危型HPV感染妇女中49例HPV L1壳蛋白表达阳性,HPV L1壳蛋白表达阳性率为20.4%。NILM组、CIN1组、CIN2组、CIN3组和SCC组HPV L1壳蛋白表达的阳性率分别为9.8%、66.7%、32.1%、9.6%和0%,五组间HPV L1壳蛋白表达阳性率之间的差异具有统计学意义(P<0.001)。通过Spearman等级相关分析,我们进一步发现HPV L1壳蛋白表达的阳性率与宫颈病变的恶性程度呈负相关(rs=-0.38,P<0.001)。结论随着宫颈病变恶性程度的增高,hTERC基因的阳性扩增率呈上升的趋势,HPVL1壳蛋白的阳性表达率呈现下降趋势,HPVL1壳蛋白与hTERC基因检测在临床上可以作为宫颈病变诊断及进一步评估进展风险的有效指标。

Objective To evaluate the significance of HPVL1 protein, hTERC gene in the cytologic specimens of cervix which was infected by the high-risk types of human papilloma virus(HR-HPV), to study expression and level of the HPVL1 protein, hTERC in correlated with cervical lesions among the people who were infected by HR-HPV. Methods 240 patients with HR-HPV positive were enrolled and the expression of HPVL1 capsid protein was detected with immunocytochemical method, and the expression of hTERC gene was detected with fluorescence chromosomal in situ hybridization(FISH), and the expression of HPVL1 protein and hTERC gene in different grades of cervical exfoliative cells were analyzed statistically. Results Genomic amplification of hTERC positive rate in NILM, CIN1, C1N2, CIN3, SCC group was 7.80%, 16.7%, 75%, 98.6%, 100%, respectively. The expression of hTERC positive rate between the five groups was statistically significant(χ2=131.9, P<0.001). The percentage of hTERC increased with the severity of the cytologic interpretation(rs=0.71, P<0.001). The positive rate of HPVL1 in NILM, CIN1, CIN2, CIN3, SCC group were 9.8%, 66.7%, 32.1%, 9.6%, 0%, respectively. The differences of the expression of HPVL1 between the five groups was statistically significant(P<0.001), the positive rate of the CIN1 group was the highest. With the increase in the degree of malignancy of cervical lesions, HPVL1 positive rate decreased, and the degree of malignancy of cervical lesions was negatively correlated(rs=-0.38, P<0.001). Conclusions hTERC expression tended to increase with cervical lesions severity; With the increase in the degree of malignancy of cervical lesions, HPVL1 positive rate decreased. HPVL1 and hTERC has an important role in shunting for treatment of the cervical intraepithelial neoplasia among who was infected by high-risk HPV.