FOXM1 Promotes Drug Resistance in Cervical Cancer Cells by Regulating ABCC5 Gene Transcription.

OBJECTIVE: The aim of the present study was to investigate the effect of forkhead box M1 (FOXM1) to paclitaxel resistance in cervical cancer cells, to determine the underlying mechanism, and to identify novel targets for the treatment of paclitaxel-resistant cervical cancer. METHODS: Paclitaxel-resistant Caski cells (Caski/Taxol cells) were established by intermittently exposing the Caski cells to gradually increasing concentrations of paclitaxel. The association between FOXM1, ATP-binding cassette subfamily C member 5 (ABCC5), and cervical cancer cell drug resistance was assessed by overexpressing or knocking down the expression of FOXM1 in Caski or Caski/Taxol cells. The protein and mRNA expression levels, the ratio of cellular apoptosis, and cell migration as well as intracellular drug concentrations were measured in cells following the different treatments. RESULTS: After the successful establishment of resistant Caski/Taxol cells, cell cycle distribution analysis showed that a significantly larger percentage of Caski/Taxol cells was in the G0/G1 stage compared with the Caski cells (P < 0.01), whereas a significantly larger percentage of Caski cells was in the S and G2/M stage compared with the Caski/Taxol cells following treatment with paclitaxel (P < 0.01). Both the protein and mRNA expression levels of FOXM1 and ABCC5 transporters were significantly higher in the paclitaxel-resistant Caski/Taxol cells compared with Caski cells (P < 0.05). Knockdown of FOXM1 significantly lowered the protein expression levels of FOXM1 and ABCC5. Intracellular paclitaxel concentrations were significantly higher amongst the Caski/Taxol cells following the knockdown of FOXM1 by shRNA or Siomycin A (P < 0.05). CONCLUSION: FOXM1 promotes drug resistance in cervical cancer cells by regulating ABCC5 gene transcription. The knockdown of FOXM1 with shRNA or Siomycin A promotes paclitaxel-induced cell death by regulating ABCC5 gene transcription.

Expression and Clinical Significance of KLRG1 and 2B4 on T Cells in the Peripheral Blood and Tumour of Patients with Cervical Cancer.

BACKGROUND: Killer cell lectin-like receptor G1 (KLRG1) and 2B4 play important roles in the immune regulation and immune tolerance to tumor cells by inhibiting T cell function. However, the clinical relevance of KLRG1 and 2B4 to cervical cancer remains to be understood. METHODS: We measured the frequency of KLRG1+ or 2B4+ cells in CD4+ or CD8 + T cells derived from peripheral blood or tumour biopsies in cervical cancer patients by flow cytometry. RESULTS: Compared with healthy controls, the level of KLRG1 and 2B4 on CD8 + T cells in the blood of the patients increased significantly (P = .0056 and .0441). KLRG1 level on CD8 + T cells and 2B4 level on CD4 + T cells in peripheral blood were significantly higher than in tumor tissues (P < .0001 and P = .0003). Higher KLRG1 level on blood-derived CD8 + T cells was observed in patients older than 54 years (P = .001) or tested to be HPV-negative (P = .026). Tumor-infiltrated CD8 + T cells demonstrated elevated KLRG1 level in patients having pelvic lymph node metastasis (P = .016). Increased 2B4 level on blood-derived CD8 + T cells was also observed in patients older than 54 years (P < .001). KLRG1 expression on both CD4 + T (P = .0158) and CD8 + T (P = .0187) cells in the peripheral blood increased after radiotherapy. CONCLUSION: KLRG1 level on T cells was related to age and HPV in patients with cervical cancer, while 2B4 level on T cells was related to age, underlying their roles in the host immune response to cervical cancer. Radiotherapy can improve the immune function of patients.

Study of 3D-printed multi-channel applicator in patients treated with brachytherapy after cervical cancer surgery / 中华放射肿瘤学杂志

Objective:To compare the dosimetric difference between 3D-printed multi-channel applicator and conventional vaginal single-channel applicator for brachytherapy, aiming to provide guidance for patients receiving brachytherapy after cervical cancer surgery.Methods:From January 2019 to November 2020, 25 cervical cancer patients complicated with VAIN Ⅲ receiving 192Ir high-dose-rate brachytherapy after cervical cancer surgery were selected. Each patient was located by CT scanning with 3D-printed multi-channel applicator and conventional vaginal single-channel applicator, and corresponding plan and evaluation were carried out. The dose volume histogram (DVH) was obtained by inverse dose optimization algorithm. The dosimetric differences of high-risk clinical target volume (HRCTV), bladder and rectum during brachytherapy were compared with those of source applicators. The optimal treatment plan was selected. Results:D 90%, D 100%, V 100% and V 150% of the plans designed by 3D-printed individual multi-channel applicator had no significant differences compared with those designed by conventional single-channel applicator (all P>0.05). The bladder and rectal D 2cm 3 designed by 3D-printed multi-channel applicator were significantly lower than those using conventional single-channel applicator, and the differences were statistically significant (both P<0.05). Conclusion:The multi-channel individual applicator target made by 3D-printing technology has good conformal property, properly protects the bladder and rectum and possesses treatment advantages over conventional single-channel applicator.

HPV16 E6-specific T cell response and HLA-A alleles are related to the prognosis of patients with cervical cancer.

BACKGROUND: T cell epitopes are polypeptide fragments presented to T cell receptors by MHC molecules encoded by human leukocyte antigen (HLA) genes after antigen-presenting cell processing, which is the basis for the study of antigen immune mechanism and multi-epitope vaccine. This study investigated T cell response to HPV16 E6 and E7 in patients with cervical squamous cell carcinoma (CSCC). Also, the HLA-A allele distribution was compared among patients and evaluated as a factor to predict prognosis in these patients. MATERIALS AND METHODS: This study recruited a total of 76 patients with International Federation of Gynaecology and Obstetrics (FIGO) stage IIB-IIIB CSCC. Mononuclear cells were isolated from the peripheral blood before any treatment and then enzyme-linked immunosorbent spot (ELISpot) assay was employed to measure the E6 and E7-specific T cell response. HLA-A alleles were typed using Sanger sequencing-based typing techniques with DNA extracted from the peripheral blood. The correlation between the T cell responses, HLA-A allele distribution and patient prognosis were analysed using the Kaplan-Meier method, univariate and multivariate Cox proportional hazard models. RESULTS: The frequency of HPV E6-specific T cell responses in patients with pelvic lymph node metastasis was lower than that in patients without metastasis (P = 0.022). The 5-year overall survival (OS) rates of patients were 87.5% for those responding to multiple overlapping peptides, 72.7% for those responding to 1-2 overlapping peptides and 47.7% for non-responders (P = 0.032). Cox regression analysis indicated that the presence of HLA*A02:07 was independently associated with worse OS (hazard ratio [HR] 3.042; 95% confidence interval [CI] 1.348-6.862; P = 0.007), while concurrent chemoradiation therapy (CCRT) was independently associated with better OS (HR 0.475; 95% CI 0.232-0.975; P = 0.042). CONCLUSION: The results of our study demonstrated that the level of HPV16 E6-specific T cell response and HLA*A02:07 were correlated with prognosis in patients with advanced CSCC.

MicroRNA-331-3p inhibits proliferation and metastasis of ovarian cancer by targeting RCC2.

INTRODUCTION: Epithelial ovarian carcinoma (EOC) is one of the most lethal gynecologic malignancies, with a poor 5-year survival rate. Numerous studies have shown that microRNAs participate in the malignant behavior of ovarian cancer cells by directly targeting multiple oncogenes or tumor suppressor genes. MATERIAL AND METHODS: Reverse transcription-PCR was used to determine the level of miR-331-3p in EOC. Cells proliferation was measured with the Cell Counting Kit-8. Cell mobility were measured by wound-healing assay. Cell migration and invasion were measured by transwell assay. Luciferase assays were used to demonstrate that RCC2 was a directed target of miR-331-3p in EOC. Western blots were used to measure the protein expression. RESULTS: We found that the expression of microRNA-331-3p (miR-331-3p) in ovarian cancer cell lines is reduced (p < 0.01), and an increase of expression of miR-331-3p in ovarian cancer cells significantly inhibits cell proliferation (p < 0.001). Transwell and wound-healing assays showed that miR-331-3p inhibits the cell motility of ovarian cancer cells (p < 0.001). Regulator of chromosome condensation 2 (RCC2) was predicted to be a novel target for miR-331-3p. Our luciferase activity assay confirmed that RCC2 is directly targeted by miR-331-3p. RCC2 was negatively regulated by miR-331-3p (p < 0.001), and overexpression of RCC2 could restore the malignant behaviors of ovarian cancer cells, which was suppressed by miR-331-3p. CONCLUSIONS: These data indicate that miR-331-3p can inhibit proliferation, migration, and invasion of ovarian cancer cells via directly targeting RCC2. Our study provides potential therapeutic targets for the treatment of ovarian cancer.

Analysis of the poor prognostic factors affecting 48 cases of cervical stump carcinoma / 中华放射肿瘤学杂志

Objective@#To explore the poor prognostic factors of patients with cervical stump carcinoma, aiming to provide certain reference for the clinical diagnosis and treatment.@*Methods@#Clinical data of 48 patients with cervical stump carcinoma admitted to the Affiliated Tumor Hospital of Xinjiang Medical University from January 1, 2005 to December 1, 2016 were retrospectively analyzed. A total of 19 patients (40%) withⅠA-ⅡA stage cervical stump carcinoma were treated with surgery+ adjuvant therapy and 29 patients (60%) in ⅡB-Ⅳ stage received radiotherapy combined with chemotherapy. The median age of onset was 51 years old. Uterine fibroids were the main cause of subtotal hysterectomy. The average time interval from subtotal hysterectomy to definite diagnosis was 10.76 years.@*Results@#The 1-, 3-, 5-year survival rate was 98%, 83% and 74%, respectively. Univariate analysis demonstrated the time interval from subtotal hysterectomy (P=0.016), tumor diameter (P=0.016), clinical stage (P=0.036), histological grade (P=0.009), lymph node metastasis (P=0.044), parametrial invasion (P=0.046), myelosuppression (P=0.013) and radical surgery (P=0.019) were the poor prognostic factors of cervical stump carcinoma.@*Conclusions@#Poor prognosis of patients with cervical stump carcinoma is correlated with tumor diameter, clinical stage, histological grade, lymph node metastasis, parametrial invasion and myelosuppression. Histological grade is an independent risk factor.

Frequency distribution of HLA alleles and haplotypes in Uyghur women with advanced squamous cell cervical cancer and relation to HPV status and clinical outcome.

PURPOSE: This study aims to investigate the association of human leukocyte antigen (HLA) alleles and haplotypes in Uyghur women with advanced squamous cell cervical cancer (SCC). METHODS: A total of 131 Uyghur patients with advanced SCC (IIb-IVa) and 91 healthy subjects from Xinjiang province were genotyped for HLA-I and II genes using Polymerase Chain Reaction Sequence Based Typing. The different frequencies of HLA alleles and haplotypes between patients and controls were compared and the correlations were analyzed between HLA distribution and HPV status and prognosis. RESULTS: (1) The frequencies of B*51:01, DRB1*07:01, DQB1*02:01, A*01:01-C*06:02, A*01:01-DRB1*07:01, C*06:02-DQB1*02:01, DRB1*07:01-DQB1*02:01 and C*06:02-DRB1*07:01-DQB1*02:01 in cancer group were higher than control group whereas the frequencies of B*44:02, B*58:01, C*05:01, DRB1*04:01, DRB1*12:01, DRB1*13:01, DQB1*02:02, DQB1*05:02, DRB1*03:01-DQB1*02:02 and DRB1*04:01-DQB1*03:02 in cancer group were lower than control group (P < 0.05). (2) The frequencies of A*01:01-C*06:02, A*01:01-DRB1*07:01, C*06:02-DQB1*02:01, DRB1*07:01-DQB1*02:01 and C*06:02-DRB1*07:01-DQB1*02:01 in HPV positive group were lower than HPV negative group, differences of which were statistically significant (P < 0.05). (3) B*44:02 and B*58:01 were associated with reduced disease-specific survival (DSS) (P = 0.010 and 0.007). (4) Multivariate Cox proportional hazard models revealed that age, International Federation of Gynaecology and Obstetrics (FIGO) stage, tumor differentiation and allele B*58:01 as independent predictors for DSS while FIGO stage and tumor differentiation as independent factors for DFS. CONCLUSIONS: In the development and progression of advanced SCC among Uyghur population, the HLA alleles and its haplotypes play an important role. B*58:01 allele may act as an independent predictor for DSS.

Knockdown of Tripartite-59 (TRIM59) Inhibits Cellular Proliferation and Migration in Human Cervical Cancer Cells.

The tripartite motif (TRIM) family of proteins is a class of highly conservative proteins that have been implicated in multiple processes. TRIM59, one member of the TRIM family, has now received recognition as a key regulator in the development and progression of human diseases. However, its role in human tumorigenesis has remained largely unknown. In this study, the effects of TRIM59 expression on cell proliferation and migration were investigated in human cervical cancer cells. The expression of TRIM59 in clinical cervical cancer tissues and cervical cancer cells was initially determined by RT-PCR and Western blot. Specific shRNA against TRIM59 was then employed to knock down the expression of TRIM59 in cervical cancer lines HeLa and SiHa. The effects of TRIM59 knockdown on cell proliferation was assessed by MTT assay and colony formation assay. Transwell assay was conducted to reveal cell migration and invasion abilities before and after TRIM59 knockdown. Our results showed that the expression of TRIM59 was significantly elevated in cervical cancers. Knockdown of TRIM59 significantly inhibited cell proliferation and colony formation as well as cell migration and invasion abilities in cervical cancer HeLa and SiHa cells. Cell cycle progression analysis showed that TRIM59-depleted cells preferred to accumulate in the S phase. These data suggest that TRIM59 is a potential target that promotes the progression of cervical cancer.

[Pathogenic bacterium and drug resistance in cervical cancer patients complicated with reproductive tract infection].

OBJECTIVE: To determine the vaginal flora distribution in cervical cancer patients and the common pathogenic bacteria as well as drug resistance, and to explore the correlation of vaginal bacterial infection and high-risk human papillomavirus (HR-HPV) infection with cervical cancer.
 METHODS: A total of 216 patients with cervical cancer served as an experimental group, and 53 patients with chronic cervicitis served as a control group. The patients' vaginal fluid in two groups was collected before the treatment for regular bacterial culture and HPV testing. The distribution and drug resistance of bacteria in two groups of vaginal secretion were observed, and the correlation of the bacterial infection and HPV infection with the cervical cancer was analyzed.
 RESULTS: The gram-negative and gram-positive bacteria accounted for 74.38% and 21.49% in the experimental group, respectively. They were mainly resistant to ampicillin and piperacillin or penicillin and erythromycin. The gram-negative and gram-positive bacteria accounted for 42.31% and 23.08% in the control group, respectively. They were mainly resistant to ampicillin and piperacillin or penicillin. HPV-positive rates in the experiment group and the control group were 60.65% and 41.51%, respectively. There were 70 patients (32.41%) and 12 patients (22.64%) with both bacterial infection and HPV-positive infection in the experiment group and the control group, respectively. However, there was no statistical difference between the 2 groups (P>0.05). 
 CONCLUSION: Escherichia coli are the main pathogen in cervical cancer and they are highly resistant to antibiotics. Bacterial infection in genital tract is not an efficient cofactor for HPV to cause the cervical cancer.

Value and safety of abdominal paraaortic lymph node dissection in the treatment of early-stage endometrial carcinoma / 医学研究生学报

Objective Systematic pelvic lymph node ( SPLN) +abdominal paraaortic lymph node ( APLN) dissection re-mains controversial in the treatment of endometrial carcinoma , especially in the early stage of the tumor .This study aims to investigate the value and safety of APLN dissection in the treatment of early-stage endometrial carcinoma . Methods We retrospectively ana-lyzed the clinical data about 109 cases of early-stage endometrial adenocarcinoma , 56 treated by SPLN dissection ( group A ) and the other 53 by SPLN+APLN dissection ( group B ) .We compared the postoperative complications , recurrence and metastasis , and progno-sis-related factors between the two groups of patients . Results No statistically significant difference was found in the incidence rate of postoperative complications between groups A and B ( 19.64% vs26.41%, P>0.05).Recurrence and metastasis were found in 12 of the 109 patients, 10 in group A and 2 in group B (17.86%vs 3.77%, P=0.019).Multivariate logistic regression analysis showed that the independent factors of recurrence and metastasis includ -ed the differentiation degree (OR=7.385, 95%CI:1.877-29.062), pathologic stage (OR=5.444, 95%CI:1.673-17.720), range of lymph mode dissection (OR=19.171, 95%CI:2.242 -163.946), and range of lesion focus (OR=12.524, 95%CI:1.186-132.280), with the range of lymph mode dissection as the greatest influencing factor on prognosis . Conclusion SPLN+APLN dissection can reduce the recurrence and metastasis and improve the prognosis of early -stage endometrial adenocarcinoma , and therefore is safe and feasible for the treatment of the tumor .

The clinical characteristic differences between thrombosis-related edema and lymphedema following radiotherapy or chemoradiotherapy for patients with cervical cancer.

Thrombosis-related edema and lymphedema are two principal types of lower extremity edema results from radiotherapy alone or chemoradiotherapy for patients with cervical cancer. To characterize differences between them, a retrospective study was performed. We collected data including age, race, body weight, FIGO stage, histology type, platelet count, haemoglobin, time of definitely diagnosis, therapeutic regimen, edema type and which leg edema firstly occurred in. Of 40 patients who were eligible for this study, 32 were diagnosed as thrombosis-related edema and 8 diagnosed as lymphedema. The differences in patient age (p = 0.004), propotion of race (p = 0.021), the latent time (p = 0.002) and the mean platelet count (p = 0.019) were statistically significant. Among 32 patients with thrombosis-related edema, 34.4% were in stage II and 53.1% in stage III, 78.1% were squamous cell carcinoma. Among 8 patients with lymphedema, 87.5% were in stage II and 62.5% were squamous cell carcinoma. The differences were not statistically significant for weight (p = 0.94), histology type (p = 0.648), edema site (p = 0.236), haemoglobin (p = 0.088) between the two grouping patients. Although the small patient cohort is a limitation, the results suggest that the patients with thrombosis-related edema may have higher proportion, lower age, shorter latent edema time and more platelet count than those with lymphedema. Also, thrombosis-related edema was likely inclined to Uigur and lymphedema to Han race. We did not find statistical differences in weight, edema site, histology type and haemoglobin between patients with thrombosis-related edema and lymphedema.