免疫组化法检测ⅠA～ⅡA期宫颈癌盆腔淋巴结微转移的研究

目的 探讨免疫组化法检测ⅠA～ⅡA期宫颈癌根治术后盆腔淋巴结内微转移及其临床意义。方法 回顾性分析2006年1月至12月中国医科大学附属第一医院妇科27例FIGO分期为ⅠA～ⅡA期宫颈癌患者的临床资料,所有患者均行子宫广泛切除术和淋巴结清扫术。术中共取518枚淋巴结,共490枚经组织学证实无转移。对盆腔淋巴结的石蜡包埋组织以抗细胞角蛋白抗体AE1/AE3标记。结果 27例490枚淋巴结中,经免疫组化检测发现,5例患者5枚淋巴结发现微转移病灶,病例总阳性率18.5%（5/27）,淋巴结总阳性率1.02%（5/490）。在有微转移的淋巴结中,髂外淋巴结组、闭孔淋巴结组的检出率分别为2.8%（3/108）和1.5%（2/132）。所切除的淋巴结数目小于20枚与大于20枚两组间微转移的检出率差异有显著性意义（P〈0.05）。结论 宫颈癌患者阴性盆腔淋巴结中存在微转移灶,随着切除淋巴结数目的增多,微转移的检出率逐渐增大,免疫组化法检测微转移淋巴结可能对指导治疗、判断预后有积极的临床意义。     

VEGF-C对宫颈癌细胞增殖和凋亡信号传导通路的影响

目的:研究VEGF-C对体外培养的宫颈癌HeLa细胞增殖和凋亡的影响;研究VEGF-C受体KDR、信号通路PI3K、MAPK在VEGF-C对宫颈癌增殖和凋亡调控中的作用。方法:应用重组人VEGF-C蛋白体外刺激宫颈癌HeLa细胞,MTT法检测细胞增殖,流式细胞仪检测细胞周期和凋亡,Western blot检测增殖与凋亡相关基因Bcl-2、CyclinD1蛋白表达;应用KDR-Ab、信号通路PI3K抑制剂LY294002、信号通路MAPK抑制剂PD98059预处理HeLa细胞,再进行VEGF-C刺激,观察上述指标的变化。结果:重组VEGF-C(50ng/μl)刺激HeLa细胞后增殖指数增加(2.13 vs 1),细胞周期S期比率增多[(64.26±0.20)%vs(30.91±0.09)%,P<0.05],细胞凋亡率降低(3.29±0.35 vs 7.44±0.55,P<0.05);Bcl-2、Cyclin D1表达增加(P<0.05)。KDR-Ab、LY294002预处理后与VEGF-C组相比增殖指数降低,细胞周期S期比率下降,凋亡指数升高,Bcl-2、Cyclin D1表达降低。PD98059预处理后,与VEGF-C组相比增殖指数降低、细胞周期S期比率下降、Bcl-2、Cyclin D1表达降低,但对VEGF-C诱导的凋亡无明显影响。结论:外源性VEGF-C作用于肿瘤细胞自身的KDR受体,激活细胞内信号传导通路MAPK途径和(或)PI3K途径诱导Cyclin D1表达,使肿瘤细胞S期加快,促进细胞周期的进程,进而促进He-La细胞增殖;通过PI3K途径诱导Bcl-2表达,抑制凋亡。     

Downregulation of miR-599 predicts poor outcome in cervical cancer patients and promotes the progression of cervical cancer

ObjectiveCervical cancer remains a leading cause of gynecological cancer-related death. In this study, we aimed to investigate the expression pattern of miR-599 and its prognostic significance in cervical cancer.Materials and methodsThe RT-qPCR analysis was used to detect the expression levels of miR-599 in cervical cancer tissues and cell lines. The association between miR-599 expression and clinical characteristics of cervical cancer patients was analyzed using the χ² test. The Kaplan–Meier analysis and multivariate Cox proportional hazards model were used to explore the prognostic significance of miR-599. Then, CCK-8 assays, transwell migration, and invasion assays were used to assess the effects of miR-599 on tumor cell proliferation, migration, and invasion of cervical cancer cells, respectively.ResultsmiR-599 expression was significantly downregulated in cervical cancer tissues and cells compared with non-cancerous tissues and HaCaT cells, respectively. Statistical analysis revealed that miR-599 expression was associated with lymph node metastasis and FIGO stage. The miR-599 expression was an independent prognostic factor for overall survival. Functionally, overexpression of miR-599 suppressed cell proliferation, migration, and invasion of cervical cancer cells, while downregulation of miR-599 had opposite effects.ConclusionmiR-599 acts as a tumor suppressor in cervical cancer that inhibiting cell proliferation, migration, and invasion of cervical cancer cells, suggesting that miR-599 may be a potential prognostic biomarker and novel targeted strategy for cervical cancer.     

Distribution of various types of low-risk human papillomavirus according to cervical cytology and histology in northern Chinese women

Objective

To describe the distribution of specific types of low-risk (LR) human papillomavirus (HPV) among a general population of northern Chinese women.

Methods

Between 2007 and 2012, 118 096 women were tested with the HPV Geno-Array Test Kit (HybriBio) at China Medical University’s Shengjing Affiliated Hospital, Shenyang, China. Among these women, 80 418 underwent cervical cytology and colposcopic examination, and 30 961 of these had a cervical biopsy. The prevalence of HPV infection among the women was analyzed according to age, and cytologic and histologic findings.

Results

CP8304 was the most common type of LR-HPV overall, and was most prevalent in the youngest age group. The overall prevalence of LR-HPV (averaged across all types) was 1.7% in women with normal cytology, 8.8% in those with atypical squamous cells of undetermined significance (ASCUS), 8.0% in those with low-grade squamous intraepithelial lesions (LSIL), and 5.8% in those with high-grade squamous intraepithelial lesions (HSIL). LR-HPV alone, without any high-risk (HR)-HPV, was most common among women with ASCUS and cervical intraepithelial neoplasia (CIN) not otherwise specified (CINNOS) together. Co-infections of LR-HPV and HR-HPV were most common among women with LSIL and CIN1.

Conclusion

These data will facilitate modeling of the cost-effectiveness of a prophylactic LR-HPV vaccination in China.     

Overexpression of gelsolin in human cervical carcinoma and its clinicopathological significance

Objectives.

Cervical carcinoma is the second most common cause of death from gynecological cancers worldwide. Knowledge of the molecular mechanisms underlying the tumorigenesis of cervical cancer cell, except human papilloma virus infection, is limited.

Methods.

A microarray was used to study the differential expression of genes in cancerous tissues to identify new molecular markers for diagnosis and prognosis. Their differential expression was confirmed with Western blotting and immunohistochemical analyses. The clinical correlations and prognostic significance of the aberrantly expressed proteins were evaluated to identify novel biomarkers of cervical cancer.

Results.

The expression of gelsolin was significantly upregulated in 78% of patients with cervical cancer, and gelsolin was selected for further study. Gelsolin expression was stronger in cervical tumor tissues than in the surrounding noncancerous tissues (P < 0.001). Gelsolin expression in the plasma of cervical cancer patients was increased 2.2-fold compared with that of healthy control subjects (P < 0.001). The levels of plasma gelsolin in the early and late stages were significantly different (P = 0.006). According to immunohistochemical analysis, increased gelsolin expression was associated with histological type and FIGO stage II. The 5-year overall survival and recurrence-free survival rates for the low-expression group (cut-off = 115) were significantly higher than those of the high-expression group. Cancer cells with reduced gelsolin expression exhibited reduced migration and proliferation.

Conclusions.

These results provide strong evidence that gelsolin plays an important role in cellular proliferation and migration in cervical cancer and suggest that gelsolin is a promising marker for cervical cancer screening and prognosis.     

Relationship between HPV typing and abnormality of G1 cell cycle regulators in cervical neoplasm

OBJECTIVE: In tumorigenesis, loss of function of the G1 pathway (p16-CDK4/cyclinD1-pRB pathway (RB pathway) and p14-MDM2-p53 pathway (p53 pathway)) is a theoretically essential event. The simultaneous analysis of all components of the RB and p53 pathway may be able to explain cervical tumorigenesis. However, there are no reports in which all components of the G1 pathway and HPV typing were examined simultaneously in cervical cancer. METHODS: We examined HPV typing and the status of the G1 pathways simultaneously by PCR-SSCP, multiplex PCR, methylation-specific PCR, and immunohistochemical techniques in cervical neoplasia. A total of 105 samples (normal, 10; cervical intraepithelial neoplasm (CIN), 42; invasive cancer (IC), 53) were included. RESULTS: Abnormality of the RB pathway tended to be more frequent in ICs (60.4%) than in CINs (31.0%) (P = 0.069). The primary target was p16 (CIN, 14.3%; IC, 43.4%; P = 0.032). Abnormality of the p53 pathway was detected in ICs (56.6%) and in CINs (40.5%) (P = 0.1494). In particular, strong expression of MDM2 was higher in ICs (32.1%) than in CINs (7.1%) (P = 0.0045). Abnormalities of the RB and p53 pathways were higher in low-risk and negative HPV than in high-risk HPV (81.3% vs 51.4%, P = 0.0657; 81.3% vs 45.9%, P = 0.0328). Seven HPV-negative cases had abnormalities in the RB or p53 pathways. CONCLUSION: In conclusion, abnormality of the G1 pathway may be one of the important mechanisms for carcinogenesis of low-risk and negative HPV cases.     

MicroRNA miR-886-5p inhibits apoptosis by down-regulating Bax expression in human cervical carcinoma cells

Objective

MicroRNA (miRNA) plays an essential role in the progression of a variety of cancers, but its role in cervical cancer progression is not well defined. We aimed to test whether special miRNAs and their target mRNAs contribute to cervical cancer progression.

Methods

The expression profiles of 1145 microRNAs in cervical squamous cell carcinomas (CSCC) and adjacent non-tumor tissues were investigated using an Illumina microRNA microarray platform. Differentially expressed miRNAs were validated by RT-PCR. Downstream target validation was performed for miR-886-5p.

Results

We found that the expression levels of seven miRNAs differed significantly between CSCC tissues and adjacent non-tumor tissues. Forced expression of one miRNA, miR-886-5p, over-expressed in CSCC tissues lowered expression of the pro-apoptotic protein Bax, reduced apoptosis and promoted cell proliferation in H8, an HPV16-immortalized human cervical squamous epithelial cell line. Knockdown of miR-886-5p increased Bax protein and apoptotic cell death in cells of the cervical squamous carcinoma cell line, SiHa.

Conclusion

MicroRNA miR-886-5p inhibits apoptosis of cervical cancer cells by down-regulating the production of Bax.     

Diagnoses and outcomes in cervical cancer screening: a population-based study

OBJECTIVE: This study was undertaken to examine routine cervical cancer screening diagnoses and outcomes on an age-specific basis in a US population. STUDY DESIGN: We conducted an observational cohort study using 1997-2002 health plan administrative and laboratory data for women enrolled at Kaiser Permanente Northwest (Portland, Ore) in 1998. RESULTS: Across all female enrollees (n=150,052), the annual rate of routine cervical cancer screening was 294.7 per 1,000, with cytologic abnormalities detected at a rate of 14.9 per 1,000. The annual incidence of cervical intraepithelial neoplasia (CIN) 1 was 1.2 per 1,000 with a rate of 1.5 per 1,000 for CIN 2/3. CIN 1 incidence peaked among women aged 20 to 24 years (5.1 per 1,000), with CIN 2/3 rates highest among those 25 to 29 years (8.1 per 1,000). From among 44,493 routine cervical smears, results were normal for 94.5%, with abnormal diagnoses of atypical squamous cells (3.3%), atypical glandular cells (0.2%), low-grade squamous intraepithelial lesion (1.2%), high-grade squamous intraepithelial lesion (0.3%), and inconclusive/inadequate (0.5%). Of women with abnormal routine smears, CIN or cancer was detected on follow-up in 19.4% of cases, 51.5% were found to have had a false-positive smear, and 29.0% incomplete follow-up as defined by published management guidelines. CONCLUSION: These are the first comprehensive age-specific estimates of routine cervical cancer screening diagnoses and outcomes to be reported within a US general healthcare setting. Overall, 5% of routinely screened women were found to have an abnormal cervical smear with an annual incidence of CIN across all female enrollees of 2.7 per 1000.     

芬太尼通过PI3 K/Akt信号通路调控宫颈癌细胞增殖迁移

目的:探究芬太尼对宫颈癌细胞增殖迁移的作用及分子机制.方法:将宫颈癌细胞系C33 A分为对照组和芬太尼组(使用不同浓度芬太尼处理),分别采用CCK-8、Transwell法、流式细胞术检测芬太尼对C33 A细胞增殖、迁移及凋亡的影响.Western blot法检测活化Caspase 3、PARP1含量,MMP-2、MM...     

Quality of life and survival in advanced cervical cancer: a Gynecologic Oncology Group study

Purpose

To determine associations between pretreatment health-related quality of life subscales with progression-free (PFS) and overall survival (OS) in advanced and recurrent cervical cancer.

Patients and Methods

Patients included those participating in Gynecologic Oncology Group advanced or recurrent cervical cancer phase III treatment trials who completed the Functional Assessment of Cancer Therapy for patients with cervical cancer (FACT-Cx) and a single-item pain scale at study entry. The FACT-Cx includes five domains: physical (PWB), emotional (EWB), social (SWB), functional well being (FWB), and cervix cancer subscale (CCS). A high quality of life (QoL) score reflects better QoL. After stratifying by protocol and adjusting for patient and disease characteristics, a Cox proportional hazards model was fitted for each subscale as a continuous variable. If statistically significant, (p < 0.05), an analysis on mean item scores (MIS) was performed.

Results

Nine-hundred-ninety-one patients were enrolled from 1997 to 2007. The majority (87%) had recurrent disease. After adjustment for covariates and predictors, only the PWB domain (better physical QoL) was associated with improved OS [HR 0.96 95% CI 0.95-0.98; p < 0.001]. When classifying patients based on the MIS of each subscale, the patients with the lowest risk of death were likely to report less compromised QoL (MIS > 3) for PWB [HR 0.44 (0.33-0.58) P < 0.001], FWB [0.49 (0.38-0.62) P < 0.001], and CCS [0.48 (0.38-0.61) P < 0.001].

Conclusion

The pretreatment patient-reported PWB as measured by the PWB subscale of the FACT-Cx, is significantly associated with survival in advanced cervical cancer trials, even after controlling for known prognostic factors.     

Role of Toll-like receptors in cervical,endometrial and ovarian cancers: A review

Objective

The Toll-like receptors (TLRs) have been implicated in inflammation, innate immunity and cancer. The goal of this paper is to review the available published research about Toll-like receptors and their roles in gynecologic malignancies.

Methods

A Medline search was conducted and published articles from the late 1990s to the present (2014) were reviewed using search phrases, Toll-like receptors and cervical, endometrial and ovarian cancers.

Results

TLR4 and TLR5 are commonly absent in normal cervix, however TLR5 expression is strong in high grade cervical dysplasia as well as invasive cancer. The expression of TLR3 and TLR4 is low in endometrial cancer. TLR2, TLR3, TLR4 and TLR5 are highly expressed in normal and neoplastic ovarian epithelium. TLR3 has been shown to have a dual function: it can contribute to tumor elimination by upregulation of interferons α and β (INF) and natural killer cell (NK) activation or it can indirectly contribute to tumor progression.

Conclusions

Inflammation is an essential element in tumorigenesis. Toll-like receptors can trigger an inflammatory response and cell survival in the tumor micro-environment. TLRs are critical immunomodulators that may play an important role in the development of gynecologic cancers. Currently TLR agonists are being investigated for a potential role as an adjuvant in the treatment of gynecologic malignancies.     

Laparoscopic staging in advanced cervical cancer: the pros and cons of an oncological concept

In this paper, the concept of laparoscopic pretreatment staging in women with advanced cervical cancer is surveyed. While a number of authors have demonstrated the potential advantages of surgical staging for optimum individual treatment planning, clear definition of the radiation field, and potential avoidance of radical hysterectomy, an additional operation including para-aortic lymphadenectomy with considerable learning curve must also be considered. In one study, the negative effect of surgical staging on the survival of patients with cervical cancer has been reported. A positive effect of surgical staging on the prognosis of patients with advanced cervical cancer has not yet been shown. In conclusion, this concept must be further evaluated in specialized centers until a clear recommendation can be made.     

Cellular response to chemotherapy and radiation in cervical cancer

Effect of irradiation alone and irradiation after 5-fluorouracil (5-FU), paclitaxel, or cisplatin (CDDP) was investigated in human cervical cell lines (CaSki, ME180, SiHa, and C33A). High-risk human papillomavirus (HPV) (+) CaSki and SiHa cells were the most resistant to CDDP, 5-FU, and radiation treatments. Radiation and CDDP and 5-FU resulted in decreased survival of HPV 16 and 18 (+) cells, whereas addition of paclitaxel to radiation treatments decreased killing. Enhanced killing of ME180 cells containing HPV39 sequences was demonstrated with chemoradiotherapy with all agents. HPV(-) C33A was more sensitive to radiation than the other cell lines, and the addition of chemotherapeutic agents did not result in significant change in cytotoxicity. Expression of survivin was inversely proportional to cell sensitivity to CDDP, 5-FU, and radiation. Constitutive AKT levels are the lowest in cell lines that are the most resistant to CDDP, 5-FU, and radiation. These data provide correlation of response to combined therapeutic modalities with HPV status of cervical cancer and expression of survivin and AKT.     

宫颈癌组织中环氧合酶-2和诱生型一氧化氮合酶的表达及其意义

目的 :研究环氧合酶 2 (COX 2 )和诱生型一氧化氮合酶(iNOS)在宫颈癌发生发展中的作用。方法 :用免疫组化方法检测 80例宫颈癌患者COX 2和iNOS的表达水平 ,并以 3 1例宫颈上皮内瘤样病变(CIN)、3 0例慢性宫颈炎症患者和 3 0例正常宫颈上皮为对照组。结果 :(1 )宫颈癌患者COX 2和iNOS表达阳性率分别为 5 8.75 %和 86.2 5 % ,高于CIN、慢性宫颈炎症患者和正常宫颈上皮组 ;(2 )宫颈癌患者COX 2和iNOS的表达水平与临床分期、组织学分型及细胞分化程度无关 (P >0 .0 5 ) ;(3 )伴淋巴结转移的宫颈癌患者COX 2的表达水平略高于不伴淋巴结转移者 ,但差异无显著性 (P >0 .0 5 ) ,而iNOS的表达水平明显高于不伴淋巴结转移者 (P <0 .0 5 ) ;(4 )肿瘤直径≥ 5cm的宫颈癌患者COX 2的阳性表达率明显高于肿瘤直径 <5cm者 (P <0 .0 5 ) ,而iNOS的阳性表达率高于肿瘤直径 <5cm者 ,但差异无显著性 (P >0 .0 5 ) ;(5 )伴宫旁浸润或脉管浸润的宫颈癌患者COX 2和iNOS表达水平明显高于不伴宫旁浸润或脉管浸润者 (P <0 .0 1 ) ;(6)宫颈癌患者COX 2和iNOS的表达之间无相关性 ,而慢性宫颈炎症患者COX 2和iNOS的表达之间有相关性。结论 :宫颈癌患者COX 2和iNOS的表达水平明显上调 ,其表达与宫颈癌的发生发展有关     

65例早期宫颈癌耐药基因产物与临床预后关系的初步探讨

目的:探讨耐药基因产物P-糖蛋白(P-gp)、谷胱甘肽-s-转移酶(GST-π)、拓扑异构酶Ⅱ(TopoⅡ)、胸苷酸合成酶(TS)在宫颈癌中的表达及临床意义.方法:应用免疫组织化学方法检测65例宫颈癌中P-gp、TopoⅡ、GST-π、TS的表达,并分析其与临床预后之间的关系.结果:P-gp的表达与肿瘤分化程度、淋巴结转移有关(P<0.05),与UICC分期、局部复发无关(P>0.05);TopoⅡ的表达与淋巴结转移、局部复发有关(P<0.05),与肿瘤分化程度、UICC分期无关(P>0.05);GST-π的表达与各个指标之间均有关(P<0.05);TS与淋巴结转移、UICC分期有关(P<0.05),与局部复发、肿瘤分化无关(P>0.05)。经多因素Cox比例风险模型分析显示:局部复发、淋巴结转移及TopoⅡ、GST-π的表达与宫颈癌患者术后生存率有关(P<0.05)。结论:联合检测宫颈癌中P-gp TopoⅡ、GST-π、TS对判断宫颈癌的预后十分必要;局部复发、淋巴结转移及TopoⅡ、GST-π的表达可作为宫颈癌患者独立的预后因素。     

新疆地区维汉妇女宫颈病变组织中Survivin与p16~(INK4a)表达的研究

目的:探讨新疆地区维汉妇女宫颈疾病及其癌变组织中Survivin蛋白与p16INK4a蛋白的表达及意义。方法:采用免疫组化SP法,检测240例新疆地区维汉妇女宫颈病变组织中Survivin蛋白和p16INK4a蛋白的表达。维汉妇女各120例,依据病理诊断结果分为4组,非瘤变组(正常及慢性炎症组)30例,CINⅠ组30例,CINⅡ～Ⅲ组30例,宫颈鳞癌组30例。结果:宫颈病变组织中Survivin与p16INK4a蛋白在维、汉族之间的表达差异无显著性(P>0.05);随着宫颈病变程度的上升,不同组别之间Survivin与p16INK4a蛋白的表达有显著差异(P<0.05),但在CINⅡ～Ⅲ组与宫颈鳞癌组之间的表达无统计学意义(P>0.05)。结论:Survivin蛋白和p16INK4a蛋白表达与宫颈疾病及其癌变的发生关系密切,呈正相关;Survivin蛋白和p16INK4a蛋白在新疆地区维汉妇女宫颈病变及其癌变中表达不存在民族差异。