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目的探讨人宫颈癌基因(HCCR)在宫颈上皮内瘤变(CIN)和宫颈鳞癌中的表达及其与HPV16感染的关系。方法选取2013年1月-2015年3月青海大学附属医院收治的HPV16阳性CIN患者44例、宫颈鳞癌患者20例,同时选取正常宫颈组织妇女20例为对照组,均采集宫颈液基细胞残液,采用多重实时PCR技术检测宫颈脱落细胞的E2和E6基因,通过E2/E6比值法评估HPV16 DNA的体内整合状态。采用免疫组化测定各组患者HCCR表达情况。结果随着病变程度加重,HPV16-DNA游离型比例逐渐下降,正常宫颈组、CINⅠ、Ⅱ、Ⅲ、宫颈癌组分别为19例(95.0%)、9例(60.0%)、7例(38.9%)、3例(27.3%)、3例(15.0%),各组间比较差异有统计学意义(P<0.05);HPV16-DNA混合型比例逐渐增加,依次为1例(5.0%)、6例(40.0%)、10例(55.6%)、6例(54.5%)、11例(55.0%),各组间比较差异有统计学意义(P<0.05)。但CIN各组及宫颈癌组差异无统计学意义(P>0.05);而HPV16-DNA整合型逐渐增加,差异有统计学意义(P<0.05);随着病变程度加重,HCCR表达逐渐增加,各组间比较差异有统计学意义(P<0.05)。在宫颈癌组,HCCR表达阳性率在游离型、混合型和整合型中的表达率呈增加趋势;但HCCR在正常组织及CINⅠ中均无表达。宫颈癌患者中,HPV16型DNA整合状态与临床期别、分化程度、淋巴结转移有关(P<0.05),与年龄无关(P>0.05)。结论 HPV感染可能与HCCR及HPV16整合状态有关,HCCR及HPV16存在状态检测,可能作为细胞学筛查的一种补充手段,以期早发现CIN及宫颈癌的高危患者。
Objective To investigate the expression of human cervical cancer gene (HCCR) in cervical intraepithelial neoplasia (CIN) and cervical squamous cell carcinoma and its relationship with HPV16 infection. Methods From January 2013 to March 2015, 44 cases of HPV16-positive CIN, 20 cases of cervical squamous cell carcinoma and 20 cases of normal cervical tissue were selected as the control group, The E2 and E6 genes in cervical exfoliated cells were detected by multiplex real-time PCR, and the in vivo integration status of HPV16 DNA was evaluated by E2 / E6 ratio method. The expression of HCCR in each group was determined by immunohistochemistry. Results The percentage of HPV16-DNA free type gradually decreased with the severity of the disease. Among the normal cervical group, CINⅠ, Ⅱ, Ⅲ, cervical cancer group were 19 cases (95.0%), 9 cases (60.0%), 7 cases (38.9% ), 3 cases (27.3%) and 3 cases (15.0%). There was significant difference between each group (P <0.05). The proportion of HPV16-DNA mixed type increased gradually (40.0%), 10 cases (55.6%), 6 cases (54.5%) and 11 cases (55.0%). There was significant difference between the groups (P <0.05). However, there was no significant difference between CIN group and cervical cancer group (P> 0.05), but HPV16-DNA integration type increased gradually (P <0.05). HCCR expression gradually increased with the severity of lesions, The difference between each group was statistically significant (P <0.05). In cervical cancer group, the positive expression rate of HCCR in free type, mixed type and integrated type showed an increasing trend; however, HCCR was not expressed in normal tissues and CINⅠ. Cervical cancer patients, HPV16 DNA integration status and clinical stage, differentiation, lymph node metastasis (P <0.05), has nothing to do with age (P> 0.05). Conclusion HPV infection may be related to the integrated status of HCCR and HPV16. HCCR and HPV16 status may be used as a supplementary means for cytological screening in order to detect early high-risk patients with CIN and cervical cancer.