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OBJECTIVES@#Cervical squamous cell carcinoma is the most common cancer in female reproductive system. This study aims to explore the effect of microRNA-9-5p (miR-9-5p) on the migration, invasion, and epithelial-mesenchymal transition (EMT) process of cervical squamous cells.@*METHODS@#Bioinformatics were used to predict the miRNAs that could bind to E-cadherin (E-cad). The Cancer Genome Atlas (TCGA) database was used to analyze and extract significantly differentially expressed miRNAs from part of cervical squamous cell carcinoma tissues and normal cervical tissues, and miR-9-5p was selected as the main research target. The translated regions (UTR) of wild-type E-cad (E-cad-WT 3'-UTR) or the 3'-UTR of mutant E-cad (E-Cad-MUT 3'-UTR) was transfected with miR-9-5p mimic normal control (NC), and miR-9-5p mimic was co-transfected human embryonic kidney cells (293T). The relationship between miR-9-5p and E-cad was detected by double luciferase assay. The expression of miR-9-5p in normal cervical epithelial cell lines (H8) and cervical squamous cell lines (C33A, siha, caski and Me180) were detected by quantitative real-time PCR. Then, the experiments were divided into groups as follows: a block control group, an overexpression control group (mimic-NC group), a miR-95p overexpression group (mimic group), an inhibitory expression control group (inhibitor-NC group), and a miR-9-5p inhibitory expression group (inhibitor group). The changes of migration ability were detected by scratch assay. Transwell invasion assay was used to analyze the changes of invasion ability, and the mRNA and protein changes of E-cad and vimentin were detected by quantitative real-time PCR and Western blotting.@*RESULTS@#MiR-9-5p had a targeting binding relationship with E-cad. Compared with the normal cervical tissue H8 cell line, the miR-9-5p was highly expressed in cervical cancer cell lines (C33A, siha, caski and Me180) (all P<0.05). The luciferase activity of E-cad-MUT was increased compared with that of E-cad-WT in miR-9-5p mimic cells (P<0.05). Compared with the blank control group, the protein and mRNA expressions of E-cad were decreased in the miR-9-5p mimic group (both P<0.05), which were increased in the miR-9-5p inhibitor group (both P<0.05). Compared with H8 cell line, the miR-9-5p was highly expressed in the cervical squamous cell lines (all P<0.05). Compared with the mimic-NC group, the distance of wound healing, the number of caski and Me180 cells invaded below the membrane, and the mRNA and protein expressions of vimentin were all increased in the miR-9-5p mimic group (all P<0.05), while the mRNA and protein of E-cad were decreased (both P<0.05). Compared with the inhibitor-NC group, the distance of wound healing, the number of caski and Me180 cells invading the membrane, and the mRNA and protein expressions of vimentin were decreased in the miR-9-5p inhibitor group (all P<0.05), but the mRNA and protein expressions of E-cad were increased (both P<0.05).@*CONCLUSIONS@#The miR-9-5p is highly expressed in cervical squamous cell carcinoma, which can increase the migration and invasion ability, and promote the EMT process of cancer cells.
Subject(s)
Humans , Female , Cell Line, Tumor , Vimentin/metabolism , Uterine Cervical Neoplasms/genetics , Epithelial-Mesenchymal Transition/genetics , MicroRNAs/metabolism , Carcinoma, Squamous Cell/genetics , Cell Movement/genetics , RNA, Messenger , Cell Proliferation/genetics , Gene Expression Regulation, NeoplasticABSTRACT
We conducted a prospective study to assess the non-inferiority of adjuvant chemotherapy alone versus adjuvant concurrent chemoradiotherapy (CCRT) as an alternative strategy for patients with early-stage (FIGO 2009 stage IB-IIA) cervical cancer having risk factors after surgery. The condition was assessed in terms of prognosis, adverse effects, and quality of life. This randomized trial involved nine centers across China. Eligible patients were randomized to receive adjuvant chemotherapy or CCRT after surgery. The primary end-point was progression-free survival (PFS). From December 2012 to December 2014, 337 patients were subjected to randomization. Final analysis included 329 patients, including 165 in the adjuvant chemotherapy group and 164 in the adjuvant CCRT group. The median follow-up was 72.1 months. The three-year PFS rates were both 91.9%, and the five-year OS was 90.6% versus 90.0% in adjuvant chemotherapy and CCRT groups, respectively. No significant differences were observed in the PFS or OS between groups. The adjusted HR for PFS was 0.854 (95% confidence interval 0.415-1.757; P = 0.667) favoring adjuvant chemotherapy, excluding the predefined non-inferiority boundary of 1.9. The chemotherapy group showed a tendency toward good quality of life. In comparison with post-operative adjuvant CCRT, adjuvant chemotherapy treatment showed non-inferior efficacy in patients with early-stage cervical cancer having pathological risk factors. Adjuvant chemotherapy alone is a favorable alternative post-operative treatment.
Subject(s)
Female , Humans , Uterine Cervical Neoplasms/drug therapy , Prospective Studies , Quality of Life , Neoplasm Staging , Chemoradiotherapy , Chemotherapy, Adjuvant/adverse effects , Adjuvants, Immunologic , Antineoplastic Combined Chemotherapy Protocols/therapeutic use , Retrospective StudiesABSTRACT
Total hysterectomy is a treatment for cervical intraepithelial neoplasia Ⅲ (CINⅢ), and its recurrence rate is very small. In recent years, with the development of colposcopy and the increase of postoperative follow-up rate, the detection rate of vaginal intraepithelial neoplasia (VAIN) and vaginal cancer after CINⅢ hysterectomy has increased significantly. In recent years, a number of studies have found that persistent human papillomavirus (HPV) infection is the most important factor leading to the recurrence of vaginal stump after CINⅢ level hysterectomy, so eliminating HPV infection can improve the patient's performance. The prognosis of persistent HPV infection is affected by many factors. The author reviewed the factors related to the clearance of HPV after CINⅢ hysterectomy. The purpose of this study is to provide a theoretical basis for improving the prognosis of patients and delaying the recurrence time after CINⅢ hysterectomy.
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Total hysterectomy is a treatment for cervical intraepithelial neoplasia Ⅲ (CIN Ⅲ),and its recurrence rate is very small.In recent years,with the development of colposcopy and the increase of postoperative follow-up rate,the detection rate of vaginal intraepithelial neoplasia (VAIN) and vaginal cancer after CIN Ⅲ hysterectomy has increased significantly.In recent years,a number of studies have found that persistent human papillomavirus (HPV) infection is the most important factor leading to the recurrence of vaginal stump after CIN Ⅲ level hysterectomy,so eliminating HPV infection can improve the patient's performance.The prognosis of persistent HPV infection is affected by many factors.The author reviewed the factors related to the clearance of HPV after CIN Ⅲ hysterectomy.The purpose of this study is to provide a theoretical basis for improving the prognosis of patients and delaying the recurrence time after CIN Ⅲ hysterectomy.
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Objective:To explore the effect of junctional plakoglobin (JUP) on the radiotherapy-resistant cervical cancer cells in vitro.Methods:Cervical cancer cell lines SiHa, HeLa and Me-180 were irradiated with 1 Gy of 60Co radioactive rays for 3 times every week to induce the radiotherapy resistance of cells which obtained the radiotherapy-resistant cell lines SiHaIR, HeLaIR and Me-180IR. The corresponding wild-type cell lines were served as control groups. Real-time fluorescence quantitative polymerase chain reaction (RT-qPCR) and Western blot were used to detect the expressions of JUP mRNA and protein in the 6 groups of cells. The cell scratch test was used to detect the cell migration ability. Results:The relative expressions of JUP mRNA in SiHa and SiHaIR groups, HeLa and HeLaIR groups and Me-180 and Me-180IR groups were the relative expressions of JUP protein were 1.74±0.06 and 0.48±0.02 ( t = 12.327, P < 0.01), 1.77±0.06 and 0.28±0.03 ( t = 14.698, P < 0.01), 2.276±0.061 and 0.780±0.011 ( t = 7.367, P < 0.01); 2.36±0.03 and 0.55±0.02 ( t = 9.245, P < 0.01), 2.13±0.02 and 0.23±0.01 ( t = 15.643, P < 0.01), 1.96±0.05 and 0.73±0.02 ( t = 5.826, P < 0.01). When culturing the cells for 12, 24, and 48 h after scratching, the migration rates in 3 groups of radiotherapy-resistant cell lines were increased compared with the corresponding wild-type cell lines, and the differences were statistically significant (all P < 0.05). Conclusion:The expressions of JUP in radiotherapy-resistant cervical cancer cell lines are lower than those in wild-type cell lines, and the migration ability of radiotherapy-resistant cells is enhanced.
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Malignant ovariangerm cell tumors (MOGCTs) is second only to epithelial tumor which often occur in young women and young women,with high malignancy and high mortality.Effective treatment is particularly important in clinical practice.The prognosis is improved for valid chemotherapy scheme foun ded in recent years.Surgery still play a crucial role in the therapy of MOGCTs no matter for the primary operation or re-operation.Since the 70s,the comprehensive surgical staging (CSS) has improved the prognosis in patients with malignant ovarian cell tumors.Retroperitoneal lymphadenectomy is an integral part of the complete staging in MOGCT.The paper discusses various aspects of the clinical value of comprehensive surgical staging in MOGCTs.
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Objective To investigate the effects of specific methyltransferase inhibitor of 5-Aza-2'-deoxycytidine (5-Aza-CdR) on the promoter methylation of E-cadherin (E-cad) gene, protein expression in human cervical cancer SiHa cells, and the cell biological behavior. Methods SiHa cells were treated with 5-Aza-CdR at different concentrations. Quantitative methylation-specific polymerase chain reaction (QMSP) was used to examine CpG island promoter methylation level of E-cad gene before and after treatment. The experimental group of the optimum concentration was selected. The expression levels of E-cad mRNA and its protein in SiHa cells line were detected by quantitative real-time polymerase chain reaction (RT-PCR) and western blot respectively. Cell adhesion test was used to measure cell adhesion ability and Transwell test was used to detect cell invasion and migration ability. Results E-cad gene promoter methylation index (PMR) of 5-Aza-CdR at 0, 1, 5, 10, 15 μmol/L level was (53.0 ±1.6) %, (50.0 ±1.2) %, (44.0 ±1.4) %, (27.0 ±1.7) %, (15.0±8.2) %respectively, and PMR value decreased gradually with the increase of 5-Aza-CdR concentration. Furthermore, PMR value was the lowest at 15μmol/L, and the difference was statistically significant compared with other 4 groups (P< 0.01). Then 5-Aza-CdR at 15 μmol/L was selected as the following experimental concentration. The expression of E-cad mRNA and its protein in the 5-Aza-CdR group were significantly higher than those in the blank control group (P<0.05). The rates of cell adhesion , cell invasion inhibition and migration inhibition were all increased with significant differences (P<0.05). Conclusions 5-Aza-CdR can upregulate E-cad mRNA and protein expression level in cervical cancer SiHa cells, reduce cell invasion and migration ability, and promote the adhesion of SiHa cells, which has reversed hypermethylation in the promoter region of E-cad gene partly.
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Objective To investigate the effects of specific methyltransferase inhibitor of 5-Aza-2'-deoxycytidine (5-Aza-CdR) on the promoter methylation of E-cadherin (E-cad) gene, protein expression in human cervical cancer SiHa cells, and the cell biological behavior. Methods SiHa cells were treated with 5-Aza-CdR at different concentrations. Quantitative methylation-specific polymerase chain reaction (QMSP) was used to examine CpG island promoter methylation level of E-cad gene before and after treatment. The experimental group of the optimum concentration was selected. The expression levels of E-cad mRNA and its protein in SiHa cells line were detected by quantitative real-time polymerase chain reaction (RT-PCR) and western blot respectively. Cell adhesion test was used to measure cell adhesion ability and Transwell test was used to detect cell invasion and migration ability. Results E-cad gene promoter methylation index (PMR) of 5-Aza-CdR at 0, 1, 5, 10, 15 μmol/L level was (53.0 ±1.6) %, (50.0 ±1.2) %, (44.0 ±1.4) %, (27.0 ±1.7) %, (15.0±8.2) %respectively, and PMR value decreased gradually with the increase of 5-Aza-CdR concentration. Furthermore, PMR value was the lowest at 15μmol/L, and the difference was statistically significant compared with other 4 groups (P< 0.01). Then 5-Aza-CdR at 15 μmol/L was selected as the following experimental concentration. The expression of E-cad mRNA and its protein in the 5-Aza-CdR group were significantly higher than those in the blank control group (P<0.05). The rates of cell adhesion , cell invasion inhibition and migration inhibition were all increased with significant differences (P<0.05). Conclusions 5-Aza-CdR can upregulate E-cad mRNA and protein expression level in cervical cancer SiHa cells, reduce cell invasion and migration ability, and promote the adhesion of SiHa cells, which has reversed hypermethylation in the promoter region of E-cad gene partly.
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Objective To investigate the influence of human papillomavirus (HPV) 16 E6 gene silencing by small interfering RNA (siRNA) on the expression and the promoter hypermethylation status of E-cadherin (E-cad) in cervical cancer SiHa cell line. Methods siRNA which used lentivirus as the vector was used to knock down the HPV16E6 gene in cervical cancer SiHa cell line. The expression levels of HPV16E6 mRNA, E-cad mRNA and protein in siRNA-HPV16E6 SiHa cell line were detected by RT-qPCR and Western blot, respectively. Methylation specific PCR (MSP) method was used to detect the methylation status of E-cad gene (CDH1) promoter in siRNA-HPV16E6 SiHa cell line. Results The E-cad mRNA expression levels in siRNA E6 group, empty vector group and blank control group were 4.755±1.085, 1.224± 0.840, 1.327±1.221, respectively. The protein expression levels were 0.616±0.019, 0.325±0.016, 0.299±0.015, respectively. The expressions of E-cad mRNA and protein in siRNA E6 group were significantly higher than those in the empty vector group and blank control group (F = 21.346, P 0.05). After knocking down HPV16E6 gene, the methylation status of E-cad gene was weakly positive, and the intensity of the amplified products was significantly weaker than that in the empty vector group and blank control group, while the unmethylation amplification was positive. Conclusions Knocking down the HPV16E6 gene increases the expression of E-cad in cervical cancer SiHa cell line, and decreases the level of CDH1 promoter methylation. To a certain extent, it partly reverses the hypermethylation status of CDH1 promoter, and causes E-cad to be re-expressed.
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Objective To investigate the expressions of human papillomavirus (HPV) 16E6 and E-cadherin in different cervical tissues.Methods Immunohistochemistry expressions of HPV16E6 and E-cadherin were analyzed in 30 normal cervical tissues,34 cervical intraepithelial neoplasias (CIN),and 60 squamous cervical cell carcinomas.Results The differences in positive expression rates of HPV16E6 and E-cadherin were statistically significant among normal cervical tissues,CIN Ⅰ,CIN Ⅱ-Ⅲ,cervical squamous cell carcinomas (x2 =40.166,P =0.000;x2 =31.295,P =0.000),respectively.The abnormal expressions of HPV16E6 and E-cadherin expression were significantly related to clinical pathological stage,tumor size,histological grade,invasion depth of squamous cervical cancers (P < 0.05),but not correlated with age (P >0.05).The expressions between HPV16E6 and E-cadherin protein were negatively correlated (rs =-0.647,P =0.000).Conclusions There may be relationship between HPV16E6 and E-cadherin,and the abnormal expressions of two proteins may be associated with poor prognosis of cervical carcinomas.
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OBJECTIVE@#To analyze the expression of vascular endothelial growth factor (VEGF), miR-205, Ezrinand Lamin A/C in ovarian cancer tissues.@*METHODS@#The expression of VEGF in the serum of epithelial ovarian cancer and that of healthy volunteers were detected by enzyme-linked immunosorbent assay; the expressions of vascular endothelial growth factor receptor 1 (VEGFR-1), VEGFR-2, Ezrin and Lamin A/C were detected by immunohistochemistry and the micro-vessel density (MVD) of CD31 was detected by immunohistochemistry in epithelial ovarian cancer, benign ovarian and normal ovarian specimens; and the expression of miR-205, Ezrin and Lamin A/C were detected by real-time PCR in epithelial ovarian cancer, benign ovarian and normal ovarian specimens.@*RESULTS@#The expression of VEGF in the serum of epithelial ovarian cancer patients (116.10± 11.94) was significantly higher than that of healthy volunteers (40.04±4.97, P0.05). The average length of MVD in the epithelial ovarian cancer specimens (7.56±0.51), was significantly higher than that in the normal ovarian specimens (1.22±0.56, P0.05). The relative expression level of miR-205 was 0.106±0.035 in the epithelial ovarian cancer specimens, which was significantly higher than that in the normal ovarian specimens (0.0007±0.0005, P0.05). The positive expression rates of Ezrin and Lamin A/C in the epithelial ovarian cancer specimens were 51.7% and 60.3%, respectively, which were significantly lower than those in the benign ovarian and the normal ovarian specimens (P0.05). The relative expression levels of Ezrin and Lamin A/C mRNA in the epithelial ovarian cancer specimens were (0.026±0.003) and (0.060±0.007), respectively, which were significantly lower than those in the normal ovarian specimens (P0.05) .@*CONCLUSION@#VEGF is significantly expressed in the serum of epithelial ovarian cancer patients; and miR-205 is up-regulated in the epithelial ovarian cancer specimens. Ezrin and Lamin A/C are down-regulated in the epithelial ovarian cancer samples. VEGF, miR-205 and target protein may be associated with the invasion and metastasis of epithelial ovarian cancer.
Subject(s)
Female , Humans , Carcinoma, Ovarian Epithelial , Cytoskeletal Proteins , Genetics , Metabolism , Down-Regulation , Enzyme-Linked Immunosorbent Assay , Immunohistochemistry , Lamin Type A , Genetics , Metabolism , MicroRNAs , Genetics , Metabolism , Neoplasms, Glandular and Epithelial , Genetics , Metabolism , Ovarian Neoplasms , Genetics , Metabolism , RNA, Messenger , Real-Time Polymerase Chain Reaction , Vascular Endothelial Growth Factor A , Blood , Vascular Endothelial Growth Factor Receptor-1 , Metabolism , Vascular Endothelial Growth Factor Receptor-2 , MetabolismABSTRACT
Objective To investigate the value of DNA ploidy analysis in cervical cancer screening for outpatients.Methods 840 from 2 692 outpatients examed by Thin Prep cytology,DNA ploidy analysis were performed directed biopsy simultaneously.Sample were taken by cervix brush and transported into a fixative solution.Two slides were made from each sample for staining with Feulgen DNA specific staining and the other with Pap stained,respectively.The routine cytological diagnosis of Pap smear was done by cytology physicians,and the Feulgen staining tablets by the automated DNA ploidy analysis system.Results Among 840 cases,554 cases (66.0 %) were histological diagnosed as chronic cervicitis,25 cases (3.0 %) as cervical intraepithelial neoplasia (CIN) Ⅰ,59 cases (7.0 %) as CIN Ⅱ,100 cases (11.9 %) as CINⅢ and 102 cases (12.1%) as cervical invasive cancer by pathological biopsy.486 cases were observed with DNA heteroploid and 354 were not.The sensitivity,specificity,positive predictive values and negative predictive values of scanning CIN Ⅱ or more severe cervical diseases by DNA heteroploid positive or heteroploid ≥3 for were 91.9 % or 89.2 %,58.5 % or 35.8 %,49.4 % or 57.3 %,94.1% or 77.2 %,respectively,while those of scanning equal or more than LSIS andthe above diseases by Thin Prep cytology were 40.2 %,90.0 %,39.6 % and 76.9 %.Conclusion DNA ploidy analysis might be a useful tool for cervical cancer screening and has a competitive sensitivity compared with conventional cytology.
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Objective To explore the security,pregnancy outcomes,and the tumor recurrence related factors of young patients with cervical cancer treated with different radical trachelectomy (RT).Methods Thirty-two young patients < 40 years of age with early cervical cancer from May 2004 to July 2012 admitted in Tumor Hospital Xiangya School of Medicine of Central South University were divided into two groups based on different operation methods:vaginal radical trachelectomy (RVT) group and abdominal radical trachelectomy (RAT) group.The clinical data were analyzed by One-way Anova and multivariate Cox stepwise regression analysis.Results The operation duration,number of lymph node dissection,the height of the cervical resection,postoperative hospitalization time,incidence of vascular injury and incidence of postoperative lymphocele were respectively (250 ± 82) min,15 ± 6,(2.31 ± 0.21) cm,(9.2 ± 2.9) d,1/18 and 1/18 in RVT group,while (263 ±60) min,16 ±8,(2.32 ±0.26) cm,(10.3 ±3.5) d,0 and 1/14 in RAT group.There was no statistically significant difference between the two groups (all P > 0.05).The blood loss (281 ±201) ml in RVT group was significantly lower than that in the RAT group (492 ±320) ml (P <0.05).The length of Vaginal hysterectomy[(2.61 ±0.50) cm] and the width of parametrial resection[(2.38 ±0.36) cm] in RVT group were significantly less than those[(2.95 ±0.10),(2.81 ±0.22) cm] in the RAT group (all P < 0.05).The pregnancy rate between RVT group (3/18) and RAT group (2/14) were no significant difference (P > 0.05).One-way Anova analysis showed that the recurrence of early cervical cancer was related to tumor size in diameter (F =4.911,P =0.047),while there were no correlation with age,clinical stage,histological type and surgical approach (all P > 0.05).Multivariate analysis showed that tumor diameter size was an independent risk factor for tumor recurrence (3 =0.259,P =0.031).Conclusions RT for young patients with early cervical cancer is feasible.Pregnancy outcomes after RT need to be study in the future.Tumor size in diameter is the major risk factor for tumor recurrence.
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Objecfive To investigate the relationship between single nucleotide polymorphism-56(SNP-56)in calpain-10(CAPN-10)gene and polycystic ovary syndrome(PCOS)in Chinese.Methods The genotypes of SNP-56 of CAPN-10 were determined through polymerase chain reaction Tm-shift genotyping method in 638 local women in Shandong Province.Among them,334 were patients with PCOS (PCOS group)and 304 were normal women(control group).The baseline parameters including levels of serum follicle-stimulating hormone(FSH),luteinizing hormone(LH),prolactin(PRL),estradiol(E2),testosterone(T)and lipid,as well as the body mass index(BMI)and waist/hip ratio(WHR)were measured.Glucose tolerance and insulin releasing before and after loading with 75 g of glucose were also assayed.Results(1)The frequencies of two allelotypes or three genotypes did not differ between PCOS women and normal women(P>0.05).(2)In PCOS group,patients with AA genotype had a significantly higher plasma glucose of 180 minutes OGTT(5.7±2.2)mmol/L[P<0.01 compared to Gagenotype(4.9±1.2) mmol/L,P<0.01 compared to GG genotype(4.9±1.4)mmol/L]and serum total cholesterol(TC)level(4.9±1.0)mmol/L[P<0.05 compared to Gagenotype(4.5±0.9)mmol/L].(3)Compared to PCOS patients with GA+GG genotype(P<0.05,P<0.01)or GG genotype(P<0.05,P<0.01),there was significantly higher attack rate of diabetes and tumor in the family history of patients with AA genotype.Conclusions These findings suggest that CAPN-10 gene SNP-56 which may not contribute to the genetic susceptibility of PCOS plays a role in glucose and lipid metabolism in Chinese PCOS patients.It may also be correlated with attack rate of diabetes and tumor in the family history of PCOS patients.
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Objective To evaluate the effect of preoperative chemotherapy on bulky cervical cancer by internal iliac arterial infusion. Methods One hundred and eighty-six patients with bulky cervical cancer were randomly divided into two groups: chemotherapy + radiotherapy group (C+R group, n=105) and radiotherapy group (R group, n=81). Patients in C + R group underwent internal iliac arterial infusion chemotherapy by using Seldinger technique internal iliac arterial or epigastric arterial catheterization. Combined regimens were prescribed including cisplatin as the major drug. Meanwhile ~192 Ir high-dose-rate intracavitary radiotherapy was performed,with A point dose at ~12-24 Gy/2-4 times (C + R group). Patients in R group were only given radiotherapy. Both groups of patients received radical hysterectomy two weeks after radiotherapy. Results The tumor regression rate of C+R group was 97.1%, significantly higher than 79.0% in R group(P0.05). Postoperative pathologic examinations showed the percentage of cervical tumor residue, parauterine invasion,pelvic lymph node metastasis in C+R group was lower than those of R group(P