Xin Wang

THUMPD3‐AS1 inhibits ovarian cancer cell apoptosis through the miR‐320d/ARF1 axis

Abstract Ovarian cancer is one of the most common gynecologic malignancies that has a poor prognosis. THUMPD3‐AS1 is an oncogenic long noncoding RNA (lncRNA) in several cancers. Moreover, miR‐320d is downregulated and inhibited proliferation in ovarian cancer cells, whereas ARF1 was upregulated and promoted the malignant progression in epithelial ovarian cancer. Nevertheless, the role of THUMPD3‐AS1 in ovarian cancer and the underlying mechanism has yet to be elucidated. Human normal ovarian epithelial cells (IOSE80) and ovarian cancer cell lines (CAVO3, A2780, SKOV3, OVCAR3, and HEY) were adopted for in vitro experiments. The functional roles of THUMPD3‐AS1 in cell viability and apoptosis were determined using CCK‐8, flow cytometry, and TUNEL assays. Western blot was performed to assess the protein levels of ARF1 , Bax , Bcl‐2 , and caspase 3 , whereas RT‐qPCR was applied to measure ARF1 mRNA, THUMPD3‐AS1 , and miR‐320d levels. The targeting relationship between miR‐320d and THUMPD3‐AS1 or ARF1 was validated with dual luciferase assay. THUMPD3‐AS1 and ARF1 were highly expressed in ovarian cancer cells, whereas miR‐320d level was lowly expressed. THUMPD3‐AS1 knockdown was able to repress cell viability and accelerate apoptosis of OVCAR3 and SKOV3 cells. Also, THUMPD3‐AS1 acted as a sponge of miR‐320d , preventing the degradation of ARF1 . MiR‐320d downregulation reversed the tumor suppressive function induced by THUMPD3‐AS1 depletion. Additionally, miR‐320d overexpression inhibited ovarian cancer cell viability and accelerated apoptosis, which was overturned by overexpression of ARF1 . THUMPD3‐AS1 inhibited ovarian cancer cell apoptosis by modulation of miR‐320d / ARF1 axis. The discoveries might provide a prospective target for ovarian cancer treatment.

Comprehensive assessment of postoperative recurrence and survival in patients with cervical cancer

The prediction of postoperative recurrence and survival in cervical cancer patients has been a major clinical challenge. The combination of clinical parameters, inflammatory markers, intravoxel incoherent motion diffusion-weighted imaging (IVIM-DWI), and MRI-derived radiomics is expected to support the prediction of recurrence-free survival (RFS), disease-free survival (DFS), tumor-specific survival (CSS), and overall survival (OS) of cervical cancer patients after surgery. A retrospective analysis of 181 cervical cancer patients with continuous follow-up was completed. The parameters of IVIM-DWI and radiomics were measured, analyzed, and screened. The LASSO regularization was used to calculate the radiomics score (Rad-score). Multivariate Cox regression analysis was used to construct nomogram models for predicting postoperative RFS, DFS, CSS, and OS in cervical cancer patients, with internal and external validation. Clinical stage, parametrial infiltration, internal irradiation, D-value, and Rad-score were independent prognostic factors for RFS; Squamous cell carcinoma antigen, internal irradiation, D-value, f-value and Rad-score were independent prognostic factors for DFS; Maximum tumor diameter, lymph node metastasis, platelets, D-value and Rad-score were independent prognostic factors for CSS; Lymph node metastasis, systemic inflammation response index, D-value and Rad-score were independent prognostic factors for OS. The AUCs of each model predicting RFS, DFS, CSS, and OS at 1, 3, and 5 years were 0.985, 0.929, 0.910 and 0.833, 0.818, 0.816 and 0.832, 0.863, 0.891 and 0.804, 0.812, 0.870, respectively. Nomograms based on clinical and imaging parameters showed high clinical value in predicting postoperative RFS, DFS, CSS, and OS of cervical cancer patients and can be used as prognostic markers.

Single-cell EMT-related transcriptional analysis revealed intra-cluster heterogeneity of tumor cell clusters in epithelial ovarian cancer ascites

Malignant ascites of epithelial ovarian cancer is a metastatic tumor microenvironment in which large amounts of disseminated single cells (DSCs) and disseminated tumor cell clusters (DTCCs) are commonly observed. The tumor cell clusters are known to be more aggressive than individual tumor cells in cancer metastasis; however, little is known about the mechanism. Applying single-cell epithelial-to-mesenchymal transition (EMT)-related transcriptional analysis in 120 DSCs and 195 intra-cluster cells from 27 DTCCs, we demonstrated that DTCCs were heterogeneous cellular units comprised of epithelial tumor cells, leukocytes, and cancer-associated fibroblasts (CAFs). Through the analysis of intra-DTCC heterogeneity, we identified that CAFs induced EMT of tumor cells via TGFβ signaling within the DTCC microenvironment. The activation of EMT program, in particular the upregulation of ZEB2, enabled the acquisition of additional chemoresistance and metastasis abilities of the intra-DTCC tumor cells, which resulted in the aggressiveness of DTCCs.

Nomogram of Combining CT-Based Body Composition Analyses and Prognostic Inflammation Score: Prediction of Survival in Advanced Epithelial Ovarian Cancer Patients

To investigate the value of body composition changes measured by quantitative computer tomography (QCT) in evaluating the prognosis of advanced epithelial ovarian cancer (AEOC) patients who underwent primary debulking surgery (PDS) and adjuvant platinum-based chemotherapy, and constructed a nomogram model for predicting survival in combination with prognostic inflammation score (PIS). Fifty-seven patients with AEOC between 2012 and 2016 were retrospectively enrolled. Pre- and post-treatment CT images were used to analyze the body composition biomarkers. The subcutaneous adipose tissue (SAT), visceral adipose tissue (VAT), cross-sectional area of paraspinal skeletal muscle area (PMA), skeletal muscle density (SMD), body mineral density (BMD) were measured from two sets of CT images. In multivariate analyses, VFA gain, PMA loss, BMD loss, and PIS were independent risk factors of overall survival (OS) (HR = 3.7, 3.0, 2.8, 1.9, respectively, all p < 0.05). Receiver operating characteristic (ROC) curves showed that the prognostic model combining body composition changes (BCC) and PIS had the highest predictive performance (area under the curve = 0.890). The concordance index (C-index) of the prognostic nomogram was 0.779 (95% CI, 0.673-0.886). Decision curve analysis (DCA) demonstrated the prognostic nomogram had a great distinguishing performance. CT-based body composition analyses and PIS were associated with poor OS for AEOC patients who underwent PDS and adjuvant platinum-based chemotherapy. The prognostic nomogram with a combination of BCC and PIS was dependable in predicting survival for AEOC patients during treatment.

Attitude and practice on human papilloma virus infection and vaccination among students from secondary occupational health school: a cross-sectional study

Cervical cancer (CC) is reported as the second-most common female cancer worldwide, of which 99% is caused by persistent human papillomavirus (HPV) infection. HPV vaccine protects against HPV infection and most cases of CC, which has only been introduced for a short time in mainland China. This study aimed to evaluate the attitude and practice related to HPV infection and vaccination among students at secondary occupational health school (SOHS) in China. We conducted a cross-sectional study in Southern China where data of 2248 participants were collected through questionnaires to estimate attitude and practice of students. Only 4.1% believed they were easily infected by HPV, 38.2% were willing to receive HPV vaccine and 30.8% intended to do regular screening of HPV infection in the future. Students in the second grade (OR = 1.51, 95%CI [1.25, 1.81]) and third grade (OR = 3.99, 95%CI [2.53, 6.27]) were more willing to take HPV vaccine compared to students in the first grade. Among the non-vaccinated participants, the most frequent reason for not receiving HPV vaccine was insufficient knowledge about HPV (91.1%). Characteristics of higher grade, personal education before enrollment and academic performance, medical specialty, history of sex experience and HPV vaccine and family history of other cancers were associated with higher attitude scores (

Plasma cells shape the mesenchymal identity of ovarian cancers through transfer of exosome-derived microRNAs

Plasma cells shape the mesenchymal identity of ovarian cancers through exosome-mediated miR-330-3p/JAM2 axis.

Single-cell RNA-seq recognized the initiator of epithelial ovarian cancer recurrence

Epithelial ovarian cancers (EOCs) are sensitive to chemotherapy but will ultimately relapse and develop drug resistance. The origin of EOC recurrence has been elusive due to intra-tumor heterogeneity. Here we performed single-cell RNA sequencing (scRNA-seq) in 13,369 cells from primary, untreated peritoneal metastasis, and relapse tumors. We used time-resolved analysis to chart the developmental sequence of cells from the metastatic tumors, then traced the earliest replanting cells back to the primary tumors. We discovered seven distinct subpopulations in primary tumors where the CYR61

Awareness and Knowledge about Human Papilloma Virus Infection among Students at Secondary Occupational Health School in China

Cervical cancer (CC) is one of the most common causes of cancer-related deaths worldwide. CC is mainly caused by human papilloma virus (HPV), which can be prevented by vaccination. We conducted a cross-sectional study in secondary occupational health school (SOHS) through a questionnaire aimed to assess the awareness and knowledge regarding HPV infection of students. A total of 2248 students participated in the survey, 45.3% of them had heard about CC, while only 21.9% of them had heard about HPV; and 74.2% had no idea of the causal link between HPV infection and CC. Most participants displayed poor awareness and knowledge about HPV infection. The results suggested that age, grade, major, academic performance, etc. were correlated with higher awareness of CC, HPV and HPV infection (p &lt; 0.05). In multivariable logistic analysis, third-grade students had the most increased awareness of CC (OR = 17.13, 95%CI: [8.11, 36.15]), HPV (OR = 6.59, 95%CI: [4.16, 10.43]) and HPV vaccine (OR = 2.78, 95%CI: [1.78, 4.32]) when compared to first-grade. Awareness and knowledge regarding HPV infection were insufficient among students in SOHS. As the future healthcare providers, these results highlight the need to supplement targeted education to improve their awareness and knowledge of HPV and vaccination.

Non-homologous dsODN increases the mutagenic effects of CRISPR-Cas9 to disrupt oncogene E7 in HPV positive cells

Genome editing tools targeting high-risk human papillomavirus (HPV) oncogene could be a promising therapeutic strategy for the treatment of HPV-related cervical cancer. We aimed to improve the editing efficiency and detect off-target effects concurrently for the clinical translation strategy by using CRISPR-Cas9 system co-transfected with 34nt non-homologous double-stranded oligodeoxynucleotide (dsODN). We firstly tested this strategy on targeting the Green Fluorescent Protein (GFP) gene, of which the expression is easily observed. Our results showed that the GFP+ cells were significantly decreased when using GFP-sgRNAs with dsODN, compared to using GFP-sgRNAs without donors. By PCR and Sanger sequencing, we verified the dsODN integration into the break sites of the GFP gene. And by amplicon sequencing, we observed that the indels% of the targeted site on the GFP gene was increased by using GFP-sgRNAs with dsODN. Next, we went on to target the HPV18 E7 oncogene by using single E7-sgRNA and multiplexed E7-sgRNAs respectively. Whenever using single sgRNA or multiplexed sgRNAs, the mRNA expression of HPV18 E7 oncogene was significantly decreased when adding E7-sgRNAs with dsODN, compared to E7-sgRNAs without donor. And the indels% of the targeted sites on the HPV18 E7 gene was markedly increased by adding dsODN with E7-sgRNAs. Finally, we performed GUIDE-Seq to verify that the integrated dsODN could serve as the marker to detect off-target effects in using single or multiplexed two sgRNAs. And we detected fewer on-target reads and off-target sites in multiplexes compared to the single sgRNAs when targeting the GFP and the HPV18 E7 genes. Together, CRISPR-Cas9 system co-transfected with 34nt dsODN concurrently improved the editing efficiency and monitored off-target effects, which might provide new insights in the treatment of HPV infections and related cervical cancer.