兰州市门诊病人人乳头瘤病毒（HPV）感染与宫颈病变相关性分析

目的：了解兰州市门诊病人人乳头瘤病毒（HPV）感染情况、年龄分布及与宫颈病变相关性,免疫组化法测不同宫颈病变组中P16表达,了解P16在筛查宫颈癌及其癌前病变中的作用。方法：收集兰州市2009年10月至2011年3月782例门诊女病人,核酸分子快速导流杂交基因芯片技术（HyBrimax）对宫颈脱落细胞行HPV分型检测,分析门诊病人中HPV感染状况及年龄分布。部分细胞学异常或高危HPV阳性者阴道镜下宫颈活检,病理检查宫颈病变程度及与HPV感染相关性,免疫组化法检测宫颈组织中P16蛋白表达情况。结果：兰州市门诊782例病人中,HPV阳性率28.64%（224/782）,21种亚型检测出17种,排在前5位亚型分别为HPV16、HPV58、HPV68、HPV52、HPV33,各年龄组间HPV感染差异有统计学意义（P〈0.05）,21-30岁之间感染占37.42%。117例病检组织中,HPV在正常/炎症、CINⅠ、CINⅡ-Ⅲ、ICC组中阳性率分别为35.55%、65.60%、88.46%、100%,各组间HPV感染差异有统计学意义（P〈0.01）,同时P16表达依次增高,阳性率分别为22.22%、56.25%、84.61%、100%,组间差异有统计学意义（X2=36.124,P=0.001）。结论：兰州市门诊病人HPV感染阳性率为28.64%,各年龄组间感染差异有统计学意义,随宫颈病变加重,HPV检出率及P16蛋白表达均增高,二者联合检测有助于早期诊断及治疗。     

SIRT1在宫颈癌细胞耐药中的作用及其机制研究

摘要目的：探讨沉默交配型信息调节因子2同源蛋白1（silentmatingtypeinformationregulation2homologyl,SIRTl）在宫颈癌化疗耐药中的作用及其机制。方法：体外培养人宫颈癌Hela细胞系和宫颈癌Hela／MMC耐药细胞亚系,westernblotting检测MMC对Hela和Hela／MMC细胞内SIRTl蛋白表达的影响;MTT法检测MMC及Nicotinamide对Hela和Hela／MMC细胞增殖的影响;AnnexinV-PI试验检测Hela／MMC细胞凋亡的亡的情况;RT—PCR方法检测耐药相关蛋白P—gP的mRNA表达情况。结果：正常情况下,Hela／MMC细胞中SIRTl的表达显著高于Hela细胞（P〈0．05）,MMC处理的Hela／MMC细胞中SIRTl的表达显著高于未经MMC处理（P〈0．05）。Nicotinamide对Hela和Hela／MMC细胞具有相似的生长抑制作用,Nicotinamide可使MMC诱导的Hela／MMC细胞凋亡增加,同时降低细胞内P-gP的mRNA表达（P〈0．05）。结论：SIRTl表达下调能显著减轻Hela／MMC细胞对MMC的耐药性,其作用可能与P—gp有关。     

Seasonality in the Incidence of Cervical Carcinoma in Teenagers and Young Adults in Northern England, 1968–2005

Infection with human papillomavirus is an established risk factor for cervical carcinoma. However, the role of other environmental factors is less well established. To further investigate whether other agents may be involved, the authors have analyzed seasonal variation in cervical cancer with respect to month of birth and separately month of diagnosis. All 85 cases diagnosed in 15–24-yr-olds during the period 1968–2005 were extracted from the specialist population-based Northern Region Young Persons' Malignant Disease Registry. The chi-square heterogeneity test was used to assess overall nonuniform variation in month of birth and separately month of diagnosis. Poisson regression analysis was used to fit sinusoidal (harmonic) models to the data using month of birth and month of diagnosis in separate models. Based on month of birth, there was statistically significant heterogeneity (p?=?.03) and a significant sinusoidal pattern, with an incidence peak involving births in the autumn months (p?=?.03). Based on month of diagnosis, there was marginally significant heterogeneity (p?=?.06). The evidence of seasonal variation around time of birth for cervical carcinoma is highly novel and suggests possible early etiological involvement of environmental factors. (Author correspondence: Richard.McNally@ncl.ac.uk)     

Synergistic antitumor activity of reversine combined with aspirin in cervical carcinoma in vitro and in vivo

A recent report showed that reversine treatment could induce murine myoblasts dedifferentiation into multipotent progenitor cells and inhibit proliferation of some tumors, and other reports showed that apoptosis of lung adenocarcinoma cells could be induced by aspirin. The aim of the present study was to evaluate the synergistic antitumor effects of reversine and aspirin on cervical cancer. The inhibition rate of reversine and aspirin on cervical cancer cell lines’ (HeLa and U14) was determined by MTT method, cell cycle of HeLa and U14 cells was analyzed by FACS, mitochondrial membrane potential of HeLa and U14 was detected using a JC-1 kit. HeLa and U14 colony formation was analyzed by soft agar colony formation assay. The expression of caspase-3, Bcl-2/Bax, cyclin D1 and p21 was detected by qRT-PCR and Western Blotting. Moreover, tumor weight and tumor volume was assessed using a murine model of cervical cancer with U14 cells subcutaneously (s.c.) administered into the neck, separately or combined with drug administration via the intraperitoneal (i.p.) route. The inhibition rate of cells in the combination group (10 μmol/L reversine, 10 mmol/L aspirin) increased significantly in comparison to that when the drugs were used alone (P < 0.05); moreover, this combination could synergistically inhibit the proliferation of five cervical cancer cell lines (HeLa, U14, Siha, Caski and C33A). In the therapeutic mouse model, tumor weight and tumor volume of cervical cancer bearing mice was more reduced when compared with the control agents (P < 0.05) in tumor-bearing mice. The combination of reversine and aspirin exerts synergistic growth inhibition and apoptosis induction on cervical cancers cells.     

TSHR蛋白在宫颈癌的表达及其与HPV-16感染的关系

目的研究促甲状腺激素受体（TSHR）在子宫颈癌组织的表达及其与乳头瘤病毒（HPV－16）的关系。方法应用免疫组织化学链霉菌抗生物素过氧化物酶（SP）法检测79例子宫颈癌和30例子宫颈炎组织HPV－16与TSHR蛋白表达。79例癌症患者中病理分级〈Ⅱ级33例,≥Ⅱ级46例;病理分期〈Ⅱ期56例,≥Ⅱ期23例;无淋巴结转移66例,有淋巴结转移13例;肿瘤大小〈3cm44例,肿瘤大小≥3cm35例。结果HPV－16在子宫颈癌表达率55．70％明显高于宫颈炎5％（P〈0．05）,TSHR在子宫颈癌表达率68．35％明显高于宫颈炎26．67％（P〈0．05）。HPV－16表达与肿瘤的大小、肿瘤分级、分期、淋巴结转移不相关。TSHR表达与肿瘤的大小呈正相关,P〈0．05,与肿瘤分级、分期及淋巴结转移不相关。HPV－16与TSHR在宫颈癌表达呈正相关。结论HPV感染对宫颈癌病变起到强烈的预警作用。TSHR不仅在甲状腺滤泡上皮细胞表达,在子宫颈癌细胞也表达,TSHR过表达能促进宫颈细胞的异常增殖,其异常功能可能是恶性肿瘤特定的临床表型。HPV与TSHR在子宫颈癌变过程中起协同作用。     

人乳头瘤病毒与宫颈癌前病变的相关性研究

目的：探讨人乳头瘤病毒感染与宫颈癌前病变两者之间的相关性。方法：选择2009年11月～2011年12月来我院就诊的220例宫颈病变患者为研究对象,将患者分成4组,即炎症组、CINl组、CIN2组和CIN3组,各组进行人乳头瘤病毒检出率及病毒载量的比较。结果：炎症组、CINl、CIN2和CIN34组人乳头瘤病毒的检出率分别为67．2％、80％、87．3％和91．4％且4组之间比较差异有显著统计学意义（P〈O．001）。炎症组HPV感染的阳性率与CIN组比较显著降低（P〈0．005）,宫颈癌前病变各组HPV感染呈阳性的病毒载量差异有统计学意义（P〈0．05）。结论：宫颈癌前病变的严重程度和感染高危型HPV病毒载量呈正相关,随着感染高危型HPV病毒载量越高,宫颈癌前病变的程度越严重。     

High TMPRSS4 expression is a predictor of poor prognosis in cervical squamous cell carcinoma

Objective The aim of this study was to clarify the clinical role of TMPRSS4 expression in cervical squamous cell carcinoma (CSCC) and to investigate the role of TMPRSS4 in predicting outcomes of patients with CSCC. Methods The retrospective study enrolled 87 patients diagnosed with CSCC between 2004 and 2006. TMPRSS4 expression in CSCC was assessed by immunohistochemistry, and data on clinical variables were collected by retrospective chart review. The impact of TMPRSS4 expression on 5-year disease-free survival (DFS) and 5-year overall survival (OS) was assessed by Kaplan–Meier analysis and Cox proportional hazards modeling. Results The high expression of TMPRSS4 was 63.2% in 87 patients with CSCC, and 17.5% in 40 patients with benign cervical disease (P < 0.001). High TMPRSS4 expression was significantly associated with tumor grade (P = 0.005), lymph node metastasis (P = 0.004), and deep cervical stromal invasion (P = 0.025). Patients with high expression of TMPRSS4 had shorter OS and DFS than those with low expression (P = 0.0205 and P = 0.0318, respectively). In multivariate Cox regression analysis, high expression of TMPRSS4 was a potential prognostic indicator for OS (P = 0.041) and DFS (P = 0.015). Conclusion Our findings suggest that TMPRSS4 might play an important role in the progression of CSCC. TMPRSS4 could be a potential prognostic marker of CSCC.     

乏氧和辐射诱导的miR-1290 促进宫颈癌细胞EMT发生

目的:研究mi R-1290在宫颈癌Hela细胞上皮间质转化中的作用。方法:模拟肿瘤放射治疗的分割疗法,单次2 Gyγ射线连续照射宫颈癌Hela、Siha细胞诱导辐射抗性细胞株;采用micro RNA芯片技术比较辐射抗性细胞与亲本细胞中mi RNA的表达谱差异;经不同条件乏氧和辐射处理宫颈癌Hela细胞后检测mi R-1290的表达水平;借助mi R-1290及mi R-1290-inhibition慢病毒表达载体,调变mi R-1290在Hela细胞的表达;调变mi R-1290后,划痕实验及Transwell侵袭实验比较Hela细胞的侵袭和转移能力;蛋白质印迹法检测细胞中E钙黏素(E-cadherin)和N钙黏素(N-cadherin)的表达。结果:在宫颈癌辐射抗性细胞中mi R-1290表达水平显著升高(P0.05);乏氧和辐射可诱导mi R-1290在宫颈癌Hela细胞中表达(P0.05);上调表达mi R-1290,Hela细胞出现了明显的间质细胞的形态改变,细胞的侵袭和转移能力明显增强(P0.05)。在Hela细胞中上调表达mi R-1290,降低了细胞中E-cadherin的表达,同时升高了N-cadherin的表达(P0.05)。结论:乏氧和辐射可诱导mi R-1290在宫颈癌Hela细胞中表达,mi R-1290通过调控E-cadherin和N-cadherin的表达促进宫颈癌Hela细胞发生EMT。     

HPV16 变异体在新疆地区宫颈癌及癌前病变中的分布研究

目的:研究新疆地区HPV16 E6、E7、LCR基因突变情况,分析HPV16变异体在宫颈癌及癌前病变发生发展中的作用。方法:选择HPV16阳性的宫颈癌及癌前病变患者,提取基因组DNA,利用PCR扩增HPV16 DNA E6、E7基因及LCR区核苷酸片段,正反向测序。与HPV16基因序列分析比对,分析核苷酸突变位点。结果:E6基因突变率为80.00%(92/115)主要突变位点T350G(59.78%)、T178G(18.47%);E7突变率为54.78%(63/115),主要突变位点A647G(33.33%)、T846C(26.98%);LCR突变率为23.48%(27/115),主要突变位点为C24T(74.07%)、C13T(25.92%)。维吾尔族T350G突变率较汉族妇女显著升高,而汉族A647G、T846C、C24T突变率显著高于维吾尔族,差异具有统计学意义(P0.05)。维吾尔族宫颈癌组T350G突变率显著高于炎症组(P0.05),且随病变严重程度增加突变率上升,汉族T350G、A647G、T846C、C24T突变率炎症组、宫颈病变组显著高于宫颈癌组(P0.05),维吾尔族C24T突变率炎症组显著高于宫颈癌组(P0.05),差异均具有统计学意义(P0.05)。结论:HPV16E6、E7突变可能与宫颈病变进展有关,T350G突变可能是维吾尔族宫颈癌高发的原因之一。