CRISPR/Cas9-based genome-wide screening for metastasis ability identifies FCGR1A regulating the metastatic process of ovarian cancer by targeting LSP1.

BACKGROUND: Metastasis is a main cause of death from ovarian cancer (OC). Identifying key markers involved in OC metastasis can aid in the effective detection of early postoperative metastasis. However, the role of FCGR1A in OC metastasis has yet to be fully established. A genome-wide CRISPR/Cas9-based screening system was used to identify regulatory factors involved in metastasis. METHODS: The expression of FCGR1A and LSP1 in ovarian cancer cell lines was examined by quantitative real-time polymerase chain reaction (qRT‒PCR). The functions of FCGR1A and LSP1 in OC cell migration, invasion and proliferation were determined using wound healing, Transwell invasion and CKK-8 assays. A transcription-activated library was used to identify the potential downstream genes of FCGR1A. FCGR1A expression was detected by immunohistochemistry and the immunity risk score (IRS) scores were calculated. RESULTS: FCGR1A was upregulated in OC cells compared with normal ovarian cells. Downregulation of FCGR1A inhibited metastasis, proliferation and epithelial-mesenchymal transition (EMT) progression in OC cells in vitro and intraperitoneal metastasis in vivo. Moreover, downregulation of FCGR1A was accompanied by decreased LSP1 expression. Overexpression of LSP1 partially reversed the tumor suppressive effect of FCGR1A downregulation. Higher FCGR1A expression was related to metastasis, higher grade, higher stage, and lymph node metastasis in OC. Survival analysis suggested that the group with higher FCGR1A expression had a lower tumor-free survival rate and a lower overall survival rate than did the group with low FCGR1A expression. CONCLUSIONS: FCGR1A enhances OC metastasis by regulating LSP1, and FCGR1A is associated with poor prognosis, suggesting that FCGR1A is a potential predictive factor for detecting early postoperative metastasis.

Different effects of CO2 laser and estrogen treatment on vaginal mucosa microbiota and function in genitourinary syndrome of menopause patients.

AIM: To characterize the effects of CO2 laser treatment and estrogen treatment on vaginal microbiota in patients with genitourinary syndrome of menopause (GSM). METHODS: Sixty-four patients with genitourinary syndrome were divided into the estrogen group, the CO2 laser group, and the control group. The control group did not receive any treatment. Vaginal mucosa was collected after 3 and 12 months of treatment. The former was used for 16S rRNA sequencing, and the latter was used for pathological evaluation. Vaginal health and voiding function were assessed using the vaginal health index (VHI) scale and the UDI-6 scale at 3 and 12 months after treatment. RESULTS: The results showed that both treatments reduced alpha diversity in the vaginal flora. Additionally, the abundance of 65 genera differed significantly between the treatment and control groups, with an increase in potentially beneficial bacteria such as Lactobacillus, IheB3_7, Mycoplasma urealyticum, and Streptococcus. In addition, the VHI and UDI-6 scores improved in both treatment groups compared to the control group after 3 months. Whereas VHI and UDI-6 scores were close to baseline in the estrogen group, and remained significantly improved in the CO2 laser group after 12 months. Pathological results showed that both methods improved the vaginal health status of patients with GSM after 12 months of treatment. However, the CO2 group exhibited a more significant increase in type III collagen. CONCLUSIONS: Both CO2 laser and estrogen therapies can regulate the vaginal flora imbalance of GSM and improve the corresponding symptoms. However, the long-term efficacy of CO2 laser therapy is superior compared to estrogen therapy.

Occurrence of hexabromocyclododecanes (HBCDs) and tetrabromobisphenol A (TBBPA) in indoor dust from different microenvironments: levels, profiles, and human exposure.

The levels and distributions of hexabromocyclododecane diastereoisomers (HBCDs) (including α, ß, and γ-HBCD) and tetrabromobisphenol A (TBBPA) were investigated in indoor dust from bedrooms and offices. HBCDs diastereoisomers were the most abundant compounds in the dust samples, and the concentrations of ∑HBCDs in the bedrooms and offices ranged from 10.6 to 290.1 ng/g and 17.6 to 1521.9 ng/g, respectively. The concentrations of target compounds in the offices were generally higher than those in the bedrooms, probably due to the presence of more electrical equipment in the offices. In this study, highest levels of target compounds were all found in the electronics. In the bedrooms, the highest mean level of ∑HBCDs was found in air conditioning filter dust (118.57 ng/g), while the personal computer table surface dust showed the peak mean concentrations of ∑HBCDs (290.74 ng/g) and TBBPA (539.69 ng/g) in the offices. Interestingly, a significantly positive correlation was observed between the concentrations of ∑HBCDs in windowsills and beddings dust in the bedrooms, suggesting beddings was one of the crucial sources of ∑HBCDs in the bedrooms. The high dust ingestion values of ∑HBCDs and TBBPA were 0.046 and 0.086 ng/kg bw/day for adults, while 0.811 and 0.04 ng/kg bw/day for toddlers, respectively. The high dermal exposure values of ∑HBCDs were 0.026 and 0.226 ng/kg bw/day for adults and toddlers, respectively. Except for dust ingestion, other human exposure pathways (such as the dermal contact with beddings and furniture) should be paid attention.

Tetrabromobisphenol A and hexabromocyclododecanes from interior and surface dust of personal computers: implications for sources and human exposure.

Tetrabromobisphenol A (TBBPA) and hexabromocyclododecane isomers (HBCDs) are widely detected in indoor environments, but the research on the accumulation, contamination, and human exposure of TBBPA and HBCDs in electronic products dust is still limited. It is unclear whether electronic products might pose human health risk via dust ingestion and dermal absorption. In this study, the levels and distributions of TBBPA and HBCDs were investigated in the personal computer (PC) interior dust and PC surface (upper and bottom) wipes. The median concentrations of TBBPA in PC interior dust, upper, and bottom surface wipes were 168.1 ng/g, 13.2 ng/m2, and 15.2 ng/m2, respectively. These levels were generally higher than those of HBCDs, which were 95.2 ng/g, 11.7 ng/m2, and 12.3 ng/m2, respectively. No significant correlations were found among the PC upper and bottom surface wipes, and interior dust, indicating different sources of TBBPA and HBCDs in PC interior and surface dust. The TBBPA and HBCDs in the PC interior dust were mainly released from inner PC materials, while the sources of target compounds on the surface wipes were likely from external environments. The exposure values of two occupational populations (including PC owners and PC repair workers) to TBBPA and HBCDs were measured by PC interior dust and upper surface wipes. The results imply dust ingestion (including hand-to-mouth uptake) is the main contributor of the exposure route to TBBPA and HBCDs for both PC owners and repair workers. Compared to PC owners, PC repair workers showed the greater risk in exposure assessment, which should be paid more attention.

A transcription factor that promotes proliferation, migration, invasion, and epithelial-mesenchymal transition of ovarian cancer cells and its possible mechanisms.

BACKGROUND: Ovarian cancer is one of the most common gynecological malignancies with the high morbidity and mortality. This study was aimed to explore the role of non-structure maintenance of chromosomes condensin I complex subunit H (NCAPH) in the progression of ovarian cancer (OC) and the transcription regulatory effects of GATA binding protein 3 (GATA3) on this gene. METHODS: Firstly, NCAPH and GATA3 expression in OC tissues and several human OC cell lines was, respectively, evaluated by TNMplot database and Western blot analysis. Then, NCAPH was silenced to assess the proliferation, migration, and invasion of OC cells in turn using CCK-8, wound healing, and transwell assays. Western blotting was used to determine the expression of epithelial--mesenchymal transition (EMT)-related proteins and PI3K/PDK1/AKT signaling proteins. The potential binding sites of GATA3 on NCAPH promoter were predicated using JASPAR database, which were verified by luciferase reporter assay and chromosomal immunoprecipitation. Subsequently, GATA3 was overexpressed to examine the biological functions of OC cells with NCAPH silencing. RESULTS: NCAPH and GATA3 expression was significantly upregulated in OC tissues and cell lines. NCAPH loss-of-function notably inhibited the proliferation, migration, invasion, and EMT of OC cells. Moreover, the expression of p-PI3K, PDK1, and p-AKT was downregulated after NCAPH knockdown. Furthermore, GATA3 was confirmed to bind to NCAPH promoter. GATA3 overexpression alleviated the inhibitory effects of NCAPH silencing on the proliferation, migration, invasion, EMT, and expression of proteins in PI3K/PDK1/AKT pathway of OC cells. CONCLUSION: To sum up, NCAPH expression transcriptional activation by GATA3 accelerates the progression of OC via upregulating PI3K/PDK1/AKT pathway.

The lncRNA TDRG1 promotes cell proliferation, migration and invasion by targeting miR-326 to regulate MAPK1 expression in cervical cancer.

BACKGROUND: Recently, lncRNA-Testis developmental related gene 1 (TDRG1) was proved to be a key modulator in reproductive organ-related cancers. The biological role of TDRG1 in cervical cancer (CC) progression remains largely unknown. METHOD: Real-time PCR (qRT-PCR) examined the expression level of TDRG1, microRNA (miR)-326 and MAPK1 mRNA. OS tissues and corresponding relative normal tissues, as well as CC cell lines and normal cell line Ect1/E6E7 were collected to determine the expression of TDRG1 in CC. MTT, colony formation, wound-healing, transwell and flow cytometer assay detected the influence of TDRG1 and miR-326 on CC cells growth, metastasis and apoptosis. Western blot examined proteins level. Bioinformatics, RNA pull-down assay, RNA immunoprecipitation and dual-luciferase reporter assays detected the molecular mechanism of TDRG1 in CC. Xenograft tumour model was established to determine the role of TDRG1 in vivo. RESULTS: The expression of TDRG1 was significantly increased in CC tissues and cell lines compared with normal tissue and normal cell line respectively and its expression was associated with clinicopathological characteristics of CC patients. Knockdown of TDRG1 inhibited the cell proliferation, migration and invasion in Hela and SIHA cells. Moreover, TDRG1 directly interacted with miR-326, and the inhibition effect on cell growth and metastasis induced by TDRG1 siRNA can be abrogated by miR-326 silencing by its inhibitor in Hela and SIHA cells. Further, MAPK1 was proved to be a direct target of miR-326, and its expression was negatively regulated by miR-326 while positively modulated by TDRG1. CONCLUSION: TDRG1 acts as a competing endogenous lncRNA (ceRNA) to modulate MAPK1 by sponging miR-326 in CC, shedding new light on TDRG1-directed diagnostics and therapeutics in CC.