Knockdown of circular RNA (CircRNA)_001896 inhibits cervical cancer proliferation and stemness in vivo and in vitro

Objective:Previous studies indicated that aberrant circular RNA(circRNA)expression affects gene expression regulatory networks,leading to the aberrant activation of tumor pathways and promoting tumor cell growth.However,the expression,clinical significance,and effects on cell propagation,invasion,and dissemination of circRNA_001896 in cervical cancer(CC)tissues remain unclear.Methods:The Gene Expression Omnibus(GEO)datasets(GSE113696 and GSE102686)were used to examine differential circRNA expression in CC and adjacent tissues.The expression of circRNA_001896 was detected in 72 CC patients usingfluorescence quantitative PCR.Correlation analysis with clinical pathological features was performed through COX multivariate and univariate analysis.The effect of circRNA_001896 downregulation on CC cell propagation was examined using the cell counting kit-8(CCK-8)test,clonogenic,3D sphere formation,and in vivo tumorigenesis assays.Results:Intersection of the GSE113696 and GSE102686 datasets revealed an increased expression of four circRNAs,including circRNA_001896,in CC tissues.Fluorescence quantitative PCR confirmed circRNA_001896 as a circular RNA.High expression of circRNA_001896 was considerably associated with lymph node metastasis,International Federation of Gynecologists and Obstetricians(FIGO)stage,tumor diameter,and survival period in CC patients.Proportional hazards model(COX)univariate and multivariate analyses revealed that circRNA_001896 expressions are a distinct risk factor affecting CC patients’prognosis.Cellular functional experiments showed that downregulating circRNA_001896 substantially suppressed CC cell growth,colony formation,and 3D sphere-forming ability.In vivo,tumorigenesis analysis in nude mice demonstrated that downregulating circRNA_001896 remarkably reduced the in vivo proliferation capacity of CC cells.Conclusion:CircRNA_001896 is highly expressed in CC tissues and is substantially related to lymph node metastasis,FIGO stage,tumor size,and survival period in patients.Moreover,downregulating circRNA_001896 significantly inhibits both in vivo and in vitro propagation of CC cells.Therefore,circRNA_001896 might be used as a biomarker for targeted therapy in cervical cancer.

Subgroups of peripheral immune effector cells in cervical cancer patients are more sensitive to radiation therapy than chemotherapy

Background:CD8 positive T lymphocytes and natural killer(NK)cells in the peripheral blood of cervical cancer patients exhibit varying sensitivities to radiotherapy and chemotherapy.Methods:A total of 50 healthy peoples and 60 cervical cancer patients were recruited.The patients with cervical cancer were separated into two groups:radiation and chemotherapy,and blood sample were collected before and after treatment.Data on the proportion of CD8 positive T lymphocytes and NK cells were gathered for analytical evaluation.Results:Compared to healthy individuals,patients with cervical cancer exhibit a reduced proportion of CD8 positive T cells within their peripheral blood.And for patients with cervical cancer,radiation therapy has been found to be more effective than chemotherapy in increasing the proportion of CD8 positive T lymphocytes and NK cells.Conclusions:These results suggest that radiation therapy increases the levels of CD8 positive T lymphocytes and NK cells within the peripheral blood of patients with cervical cancer.The study hypothesis that the changes in the percentage of CD8 positive T lymphocytes may serve as a potential indicator for predicting treatment efficacy.

Relationship between Circulating Plasma Galectin-3 Levels and T-Cell Activation during Cervical Cancer Chemotherapy

Objective: Despite the existence of several therapeutic strategies, the management of cervical cancer remains challenging. Our region has very little data on the interaction between the immune system and the clinical response to chemotherapy. This work examines plasma levels of galectin-3 (Gal-3) and percentages of activated T cells in patients with cervical cancer treated with chemotherapy and investigates if there is a relationship between the rates of these two elements. Methods: We compared data from 37 patients with cervical cancer undergoing chemotherapy and 42 controls with normal cervical cytology. Plasma Gal-3 concentrations were assessed by ELISA and expression of activation markers by T cells (CD69 and HLA-DR) was assessed by flow cytometry at three different time points during chemotherapy. Results: Our results showed that patients had a significantly higher concentration of Gal-3 compared to controls (4.025 vs. 1.340, p 0.001), similarly, they had a significantly high percentage of activated lymphocytes (2.610 vs. 0.731;p 0.0001). According to the response to treatment, patients with no response to treatment had a lower concentration of circulating Gal-3 but had approximately the same percentage of activated CD4 and CD8 lymphocytes as patients with a partial or total response. In addition, we found a positive correlation between the Gal-3 level and CD4 T cells expressing the activation marker CD69 (p 0.05;rho = 0.44). Conclusion: In conclusion, our results show that there would be a relationship between circulating galectin-3 and the percentage of peripheral CD4+CD69+ cells in cervical cancer.

EXPRESSION OF c-erbB-2 AND PCNA IN CERVICAL ADENOCARCINOMA AND ITS SIGNIFICATION

Objective: To investigate the significance of cerbB2 and PCNA expression in adenocarcinoma. Methods: Expression of cerbB2 and PCNA in 74 cases of adenocarcinoma of the uterine cervix was examined by immunohistochemistry. Results: The expression of cerbB2 was detected in 34 cases. The positive staining of cerbB2 was associated with increased lymph node metastasis (57.1% Vs 24.0%, P=0.041) and lower 5year survival rate (32.4% Vs 58.9%, P=0.008). The average PCNA labeling index (PCNA LI) was 40.6% (0.1% 91.4%). High PCNA LI was associated with lymph node metastasis (56.4% Vs 38.5%, P=0.016) and lower 5year survival rate (28.7% Vs 64.4%, P=0.005). Positive staining of cerbB2 were associated with high PCNA LI (44.7% Vs 34.6% P=0.003). Conclusions: cerbB2 and PCNA LI were associated with malignant biological behavior and poor prognosis of patients with cervical adenocarcinoma.

Treatment of uterine fibroid with triptorelin before hysterectomy

OBJECTIVE: To study the effects of pretreatment with triptorelin on uterine fibroid before abdominal hysterectomy. METHODS: Fifteen premenopausal Chinese women with symptomatic uterine fibroids requiring hysterectomy were recruited in the study. All patients received monthly intramuscular injections of 3.75 mg triptorelin for three months prior to abdominal hysterectomy. RESULTS: There was significant reduction in the serum levels of oestradiol (68.6%), progesterone (95.6%) and luteinizing hormone (73.9%) and in uterine (45.0%) and fibroid (68.0%) volumes. The serum level of follicle-stimulating hormone and haemoglobin concentration were not significantly different. CONCLUSIONS: Shrinkage of uterine fibroids can be achieved in women who are rendered hypoestrogenic with monthly injections of triptorelin for three months. This treatment modality may be of value prior to hysterectomy or myomectomy especially when the fibroid is large.

The delay, symptoms, and survival of Ivorian adolescent girls and young adults with uterine cervical cancer

Objective:To generate data on the nature and duration of cervical cancer symptoms,risk factors for delayed consultation,and diagnosis of adolescent and young adult groups in Cote d'Ivoire.Methods:This is a hospital-based cross-sectional study,conducted from July 2012 to May 2018,at the Department of Gynecology of the Yopougon teaching hospital in Cote d’Ivoire.The inclusion criteria were those who were under 40 years of age at the time of diagnosis with oral and written consent.Those whose information was more than 10%insufficient for the standards were excluded from the study.The survey files standardized anonymous pre-established data collected using medical records followed by phone calls.Results:The average age of the participants was 34±4.95 years with a minimum of 21 years.The mean parity per woman was 3.49±3.54.34.9%were HIV positive.The median patient delay was 122 days with a long delay in 84.62%of patients.The median time to total diagnosis was 209 days with a longer total delay in diagnosis of 87.18%.41%of patients honored their treatment,68.75%of which had received surgery,and 31.25%of which had received chemotherapy.The 5-year survival rate was 65%.Conclusion:The Ivorian health system must focus on primary and secondary prevention,the only guarantee to decrease the morbidity and mortality indicators linked to cervical cancer.

Evaluating the feasibility and safety of vaginal myomectomy in China

Background Classically, myomectomy has been performed via laparotomy, but laparoscopic myomectomy has now become a valuable treatment option. Vaginal myomectomy as a minimal invasive procedure has also been evaluated; however, its feasibility and safety are controversial with few clinical trials published. This study was designed to evaluate the feasibility of vaginal myomectomy in China and to document any associated complications. Methods From January 2005 to December 2010, 43 patients with symptomatic myomas were admitted for vaginal myomectomy in Peking Union Medical College Hospital （PUMCH）. The indications, operative performance, postoperative complications and outcome of these patients were analyzed retrospectively. Results Myomectomy was performed vaginally in all patients. The mean operating time was （66.4±22.6） minutes and the mean operative blood loss and hospital stay were （78.3±64.4） ml and （4.9±3.3） days, respectively. Five （12%） patients developed febrile morbidity and experienced a high postoperative temperature （mean 38.4℃）. Postoperative fever was associated with greater operative trauma, longer operative time and greater weight of the tumor （all P 〈0.01）. The cost of surgery was RMB （820.6±339.1） Yuan （（124.3±51.4） US dollars） and the total medical cost was RMB （4880.4±1088.4） Yuan （（739.5±164.9） US dollars）. Three patients later conceived spontaneously and had uneventful vaginal deliveries. Only one patient had a tumor recurrence during the following-up period. Conclusion Vaginal myomectomy is a feasible and safe surgical procedure with low recurrence and complication rates.

Feasibility and safety of vaginal myomectomy:analysis of 90 cases

Background Vaginal myomectomy was firstly reported in 1994, however, it is a relatively new technique in China. The feasibility and safety of the procedure is still controversial in this country. The aim of this study was to analyze the outcomes of vaginal myomectomy in 90 patients and to investigate the feasibility and safety of the surgery. Methods From June 2001 to June 2004, 90 patients with uterine leiomyoma were treated with vaginal myomectomy in our hospital （vaginal group）. The indications, operative performance, postoperative complications, and recovery of the patients were analyzed and compared with those of 93 patients with uterine leiomyoma treated by laparotomic myomectomy from January 2000 to January 2001 （laparotomy group）. The Student＇s t test was used to compare the continuous variables between the two groups, and the chi-square test was used to compare the categorical variables. The vaginal and laparotomy groups were followed up for 10-34 months （median, 21） and 15-24 months （median, 30）, respectively. Results Vaginal myomectomy was performed successfully in 87 of the 90 patients （96.7%）. In the other 3 patients, the procedure failed and laparotomy was carded out. In both groups, the uterus was enlarged to 8-16 weeks gestational size （median, 10 gestational weeks）. The number of resected tumors was 1 - 12 （median, 2） in the vaginal group and 1-15 （median, 4） in the laparotomy group, respectively （P〉0.05）. The mean operating time, intraoperative blood loss, and postoperative hospital stay were （52± 21） minutes, （230± 44） ml, and （7.0±1.2） days in the vaginal group, and （654±32） minutes, （200±56） ml, and （7.04±1.5） days in the laparotomy group （P〉0.05）. The mean top postoperative temperature was （38.44±1.1）℃ and （37.8±0.6）℃ in the two groups respectively （P〈0.05）. Both groups had one recurrent case during the follow-up （P〉0.05）. Condusions Vaginal myomectomy is feasible and safe in treating uterine leiomyoma. To some extent, it is superior to laparotomic myomectomy by avoiding severe trauma during the surgery.

Reconsider the safety of laparoscopic surgery in endometrial cancer

General introduction The 20th century witmessed the development of laparoscopic surgical technologies and its successful applications in gynecological and general surgeries.In 1901,Russian doctor Dimitri Ott,for the first time,inspected the human a bdominal cavity with a speculum through a small incision,which marked the origin of the concept of minimally invasive techniques."During the following decades,clinical diagnosis of intra-abdominal lesions was performed by some doctors in Europe and the US.with instuments similar to modern laparoscopy.But it was until 1980s that modem sense laparoscopic surgeries,induding laparoscopic appendicectomy and laparoscopic cholecystectomy,were conducted.In 1989,Dr.Reich Harry performned the first laparoscopic hysterectomy,'which is regarded as a comerstone in the history of gynecologic surgery.Afterwards,laparoscopic surgery rapidly stepped into the mainstream of gynecology practice during the 1990s,delivered for both benign and malignant diseases.Starting from the 2000s,the master-slave mode ro-botic system,exemplified by the da Vinci@robot,has been accepted wordwide,and currently,robot assisted laparoscopic approaches have taken up a major proportion of general,urologic and gynecologic sur-geries,especially in developed countries.5 From a historical perspective,the broad application of laparoscopic surgeries nowadays stands on solid foundations established in decades of technical revolutions and refinements.

Prevalence of high-risk human papillomavirus infection in different cervical lesion among organized health-examination women in Shanghai, China

Background Cervical cancer is the second most common cause of death from cancer among women worldwide. Human papillomavirus （HPV） plays a central role in the etiology of cervical cancer. It is important to describe the prevalence of HPV infection in different types of cervical lesions and to explore the relation between HPV viral load and the severity of cervical lesions. Methods To describe the HPV infection prevalence and viral load in different age groups, we retrospectively investigated 6405 cases of women who were organized by their units to take health-examination. They were given Hybrid Capture II tests between January 2005 and December 2006. The correlation between HPV viral load and pathology was assessed. Results Overall HPV infection prevalence was 29.1% （1864/6405）, while in women 18-20 years old it was 54.4% （31/57）, the highest among all age groups. After declining rapidly, HPV prevalence stabilized at about 30.0% in women aged 30 and older. Of the 6405 women, 1483 women had a colposcopic biopsy and 33.2% （492/1483） were positive for HPV DNA. Twenty-one percent of women with a normal diagnosis （238/1095） had HPV infection, a statistically significantly lower prevalence than in women with cervical lesions, including those with cervical intraepithelial neoplasia （68.8% in CIN1,66.7% in CIN2, and 76.5% in CIN3） or with cervical cancer （94.1%）. The correlation coefficient between viral load and cervical lesion severity was 0.134, which was not statistically significant （P=0.075）. Viral load values in women with CINs and cervical cancer were calculated, and no significant differences were identified. Conclusions The prevalence of high-risk HPV infection among women attending hospitals for health-examination in Shanghai is similar to the worldwide rate. HPV viral load can distinguish cervical lesions from normal individuals but cannot adequately predict the severity of cervical lesions.

Overexpression of Bcl-2 partly inhibits apoptosis of human cervical cancer SiHa cells induced by arsenic trioxide

OBJECTIVE: To study the biological effect of arsenic trioxide (As2O3) on human cervical cancer SiHa cells and SiHa cells overexpressing bcl-2 gene. METHODS: SiHa cells with overexpression of Bcl-2 (SiHa-Bcl2 cells) were established by transfecting SiHa cells with Bcl-2 expression vector. The sensitivities of SiHa and SiHa-Bcl2 cells to As2O3 were determined using MTT (Thiazolyl blue) reduction and colony forming ability assay, morphological analysis, flow cytometric analysis, DNA agarose gel electrophoresis, in situ cell death detection (TUNEL), Northern blot, RT-PCR and Western blot. RESULTS: As2O3 inhibited the growth of SiHa cells and induced G2/M arrest and apoptosis of the cells. RT-PCR and Western blot analysis revealed that As2O3 induced SiHa cell apoptosis possibly via inhibiting the expression of HPV16 E7 and decreasing the expression of c-myc. However, we found that SiHa-Bcl2 cells partly resisted As2O3 induced apoptosis, which might be related to the prevention of the down-regulation of HPV16 E7 and c-myc gene expression. Nevertheless, As2O3 at a high concentration could still induce apoptosis of SiHa-Bcl2 cells mainly via decreasing Bcl-2 expression and slightly inhibiting viral gene expression. CONCLUSION: As2O3 is an inducer of the apoptosis of human cervical carcinoma cells and the cells overexpressing Bcl-2 can partly resist As2O3 induced apoptosis, but the exact mechanism is unclear.

Identification of biomarkers for cervical cancer in peripheral blood lymphocytes using oligonucleotide microarrays

Background Oligonucleotide microarrays are increasingly being used to identify gene expression profiles that associated with complex genetic diseases. Peripheral lymphocytes communicate with cells and extracellular matrixes in almost all tissues and organs in human body, suggesting that the gene expression profiles in peripheral lymphocytes may reflect the presence of disease in the body. This study aimed to identify molecular biomarkers for cervical cancer in peripheral blood lymphocytes by using oligonucleotide microarrays. Methods Total RNA was extracted from peripheral blood lymphocytes of 24 early stage cervical cancer patients and 18 healthy controls. We used 22K Human Genome microarrays to profile peripheral blood lymphocytes from 4 early stage cervical cancer patients and compared their gene expression profiles with those from 3 healthy controls. Differentially expressed genes would be identified if they had adjusted P values of less than 0.05 and a groupwise average fold change greater than 1.5 or less than 0.67. Then the selected 5 genes were validated in the remaining 20 early stage cervical cancer patients and the 15 healthy controls by using real-time reverse-transcription polymerase chain reaction （RT-PCR）. Results Genes identified by the gene selection program expressed differently between the blood samples of the early stage cervical cancer patients and those of the healthy controls. To validate the gene expression data, 5 genes were analyzed by real-time RT-PCR. In three of the 5 identified genes, tenasin-c （TNC）, nuceolin （NCL）, and enolase 2 （EN02） showed a significant up-regulation in the blood samples of the early stage cervical cancer patients versus that of the healthy controls. Conclusions The up-regulation of TNC, NCL, and EN02 in peripheral blood may be used to identify novel blood biomarkers for detecting cervical cancer in a clinically accessible surrogate tissue, and thus to provide a possibility to develop a noninvasive and predictive diagnosis for the disease.

Age-specific effectiveness of primary human papillomavirus screening versus cytology in a cervical cancer screening program: a nationwide cross-sectional study

Background:Primary human papillomavirus(HPV)screening is recommended for the detection of cervical intraepithelial neoplasia(CIN)in the general pop-ulation;however,the triage for HPV-positive women remains a challenge.This study aimed to evaluate the age-specific effectiveness of primary HPV screening versus primary cytology screening for identifying optimal strategies for women of different ages.Methods:The dataset of the prevalence round screening was derived from the National Cervical Cancer Screening Program in China.Primary cervical screen-ing protocols included cytology only,HPV testing with cytology triage,and HPV testing with HPV-16/18 genotyping plus cytology triage.The primary outcomes were age-specific detection rate,colposcopy referral rate and positive predictive value(PPV)for CIN2+.Multivariate Poisson regression was used to evaluate the relativeeffectivenessofHPVtestingandcytologyaccordingtoagegroups.TheI 2 statisticwitharandom-effectmodelwasusedtotesttheheterogeneityinrelative effectiveness of HPV testing versus cytology between age groups.Results:This study included 1,160,981 women.HPV testing with HPV-16/18 genotyping plus cytology triage significantly increased the CIN2+detection by 36%(rate ratio[RR]:1.36,95%confidential interval[CI]1.21–1.54)for women aged 35-44 years and by 34%(RR:1.34,95%CI 1.20-1.51)for women aged 45-54 years compared with cytology only.HPV testing with cytology triage had simi-lar CIN2+detection rate compared with cytology only.The PPVs were substan-tially increased for both HPV testing groups.Among women aged 55-64 years old,HPV testing with HPV-16/18 genotyping plus cytology triage increased the colposcopyreferralrateby19%(RR1.19,95%CI1.10-1.29)comparedwithcytology only,butdidnotincreasetheCIN2+detection(1.09,0.91–1.30).Theeffectiveness ofHPVtestingwithcytologytriagedidnotchangeinolderwomen.Thebetween-age-group heterogeneity in the effectiveness was statistically significant for HPV testing with HPV-16/18 genotyping plus cytology triage versus cytology only.Conclusions:Our results suggested that the effectiveness of primary HPV screeningwithdifferenttriagestrategiesdifferedamongagegroups.HPVtesting with HPV-16/18 genotyping plus cytology triage could be used for women aged 35-54 years to detect more lesions,and HPV testing with cytology triage could balance the CIN2+detection and the number of colposcopies for women aged 55-64 years.Longitudinal data including both prevalence and incidence screen-ing rounds are warranted to assess age-specific triage strategies.

Immunomodulatory effects of astragalus polysaccharide on Human peripheral blood mononuclear cells co-cultured with cervical cancer cell lineImmunomodulatory effects of astragalus polysaccharide on Human peripheral blood mononuclear cells co-cultured with cervical cancer cell line

OBJECTIVE:To investigate immunomodulatory effects of Astragalus polysaccharides(APS)on the co-culture of peripheral blood mononuclear cells(PBMCs)with He La cervical cancer cell line.METHODS:To assess the proliferation of PBMCs,carboxyfluorescein succinimidyl ester(CFSE)-labeled PBMCs were co-cultured with He La cells and treated with different concentrations of APS.Supernatants of cell culture were collected for cytokines assay via enzyme-linked immunosorbent assay(ELISA).The impact of APS on the proliferation of PBMCs,induction of regulatory T cells(Tregs),and myeloid-derived suppressor cells(MDSCs)was carried out by flow cytometry.RESULTS:It was observed that APS could increase the proliferation of PBMCs co-cultured with He La cells(P<0.05).However,APS had no significant effects on the induction of Tregs and MDSCs in the co-culture assay(P>0.05).Furthermore,ELISA results demonstrated that APS could decrease IL-10 and TGF-βconcentration(P<0.05).CONCLUSION:The above-mentioned characteristics showed that APS might be able to modulate immune responses and improve anti-tumor effects through increasing the proliferation of PBMCs and decreasing inhibitory cytokines secretion as critical mediators of immune suppression in the tumor microenvironment.