宫颈鳞癌组织中miR-590-3P、FoxM1及Cdc25B细胞核内蛋白水平与高危型HPV感染的相关性研究

Correlation between miR-590-3P,FoxM1 and Cdc25B Nuclear Protein Levels and High-risk HPV Infection in Cervical Squamous Cell Carcinoma

目的:研究宫颈鳞癌组织中微小RNA-590-3P(miR-590-3P)、叉头盆蛋白质M1(FoxM1)及细胞分裂周期因子25B(Cdc25B)细胞核内蛋白水平与高危型人乳头状瘤病毒(HPV)感染的相关性。方法:将2014年2月—2019年12月医院通过宫颈活检、锥切以及子宫切除获取的宫颈组织标本300份纳入研究。将其分为正常组织59例,宫颈上皮内瘤变(CIN)1组织62例,CIN2/3组织101例,宫颈鳞癌组织78例。采用实时荧光定量PCR检测不同组织中的miR-590-3P表达水平,以免疫组织化学法检测FoxM1、Cdc25B蛋白表达情况,并进行23种基因型HPV DNA的检测。比较不同宫颈组织中的miR-590-3P以及FoxM1、Cdc25B蛋白表达情况,高危型HPV感染情况,并以Spearman相关性分析明确宫颈鳞癌组织中的miR-590-3P以及FoxM1、Cdc25B蛋白表达情况与高危型HPV感染的关系。采用受试者工作特征(ROC)曲线明确miR-590-3P以及FoxM1、Cdc25B蛋白诊断CIN2+的效能。结果:CIN2/3组织以及宫颈鳞癌组织中的miR-590-3P表达水平均高于正常组织、CIN1组织,且CIN1组织中的miR-590-3P表达水平均高于正常组织(F=46.873,P=0.000);CIN2/3组织以及宫颈鳞癌组织中的FoxM1蛋白阳性率及Cdc25B蛋白阳性率均高于正常组织及CIN1组织,且CIN1组织中的FoxM1、Cdc25B蛋白阳性率均高于正常组织(χ^(2)=47.293、51.041,P=0.000、0.000)。CIN2/3组织以及宫颈鳞癌组织中的高危型HPV感染率明显高于正常组织、CIN1组织,且CIN1组织中的高危型HPV感染率明显高于正常组织(χ^(2)=67.293,P=0.000)。经Spearman相关性分析可得:宫颈鳞癌组织中miR-590-3、FoxM1、Cdc25B蛋白表达率均和高危型HPV感染呈正相关。经ROC曲线分析发现:miR-590-3P以及FoxM1、Cdc25B蛋白联合检测诊断CIN2+的曲线下面积、灵敏度、特异度分别为0.884、0.90、0.88,均高于上述三项指标单独检测。结论:宫颈鳞癌组织中miR-590-3P、FoxM1及Cdc25B细胞核内蛋白水平与高危型HPV感染密切相关,有望成为早期诊断CIN2+的潜在生物学指标,值得临床重点关注。

Objective:To investigate the correlation between miR-590-3P,FoxM1 and cell division cycle factor 25B(Cdc25B)nuclear protein levels and high-risk human papillomavirus(HPV)infection in cervical squamous cell carcinoma.Methods:From February 2014 to December 2019,300 cervical tissue samples were collected from hospitals through cervical biopsy,cone-cutting and hysterectomy.According to the difference of disease types,59 cases of normal tissue,62 cases of cervical intraepithelial neoplasia(CIN),101 cases of CIN2/3 of the tissue,and 78 cases of cervical squamous cell carcinoma were classified.The expression levels of miR-590-3P in different tissues were detected by real-time fluorescence quantitative PCR,FoxM1 and Cdc25B protein expressions were detected by immunohistochemical method,and 23 genotypes of HPV DNA were detected.The expressions of miR-590-3P,FoxM1 and Cdc25B in different cervical tissues and high-risk HPV infection were compared.Spearman correlation analysis was used to determine the relationship between the expressions of miR-590-3P,FoxM1 and Cdc25B in cervical squamous cell carcinoma tissues and high-risk HPV infection.The efficacy of miR-590-3P and FoxM1 and Cdc25B in the diagnosis of CIN2+was determined by ROC curves.Results:The expression levels of miR-590-3p in CIN2/3 tissues and cervical squamous cell carcinoma tissues were higher than those in normal tissues and CIN1 tissues.The expression level of miR-590-3p in CIN1 tissues was higher than that in normal tissues(F=46.873,P=0.000).FoxM1 and Cdc25B protein positive rates in CIN2/3 tissues and cervical squamous cell carcinoma tissues were higher than that in normal tissues and CIN1 tissues(all P=0.000).The infection rate of high-risk HPV in CIN2/3 tissues and cervical squamous cell carcinoma tissues were significantly higher than that in normal tissues and CIN1 tissues,and the infection rate of high-risk HPV in CIN1 tissues was significantly higher than that in normal tissues(χ^(2)=67.293,P=0.000).Spearman correlation analysis showed that the expression rates of miR-590-3,FOXM1 and Cdc25b in cervical squamous cell carcinoma tissues were all positively correlated with high-risk HPV infection.ROC curve analysis showed that the area under the curve,sensitivity and specificity of the combined detection of miR-590-3p,FOXM1 and Cdc25b for the diagnosis of CIN2+were 0.884,0.90 and 0.88,respectively,which were all higher than those of the above three indicators detected alone.Conclusion:miR-590-3P,FoxM1 and Cdc25B intracellular protein levels in cervical squamous cell carcinoma are closely related to high-risk HPV infection,and are expected to be potential biological indicators for early diagnosis of CIN2+,which are worthy of clinical attention.