Ability Of Cervical Cancer Cells Research Articles

TGIF2 promotes cervical cancer metastasis by negatively regulating FCMR.

We aimed at studying the correlation between TGIF2 expression and clinicopathological features of cervical cancer (CCa). The relationship between TGIF2 and FCMR and its influence on the proliferation and metastasis of tumor cells were investigated using molecular biology techniques, so as to reveal the pathogenesis of CCa and provide a new target for clinical treatment. TGIF2 expression in 60 pairs of cervical tumors and paracancerous tissues samples collected from CCa patients of our hospital was studied by quantitative real-time polymerase chain reaction (qPCR) analysis, and the association between TGIF2 expression and the clinical indicators or prognosis of CCa patients were analyzed. CCa cells with TGIF2 knockdown were constructed using transfection technology. Changes in the biological phenotypes (proliferation, migration, invasion) of CCa cells C33-A and HeLa after TGIF2 knockdown were determined by Cell Counting Kit-8 (CCK-8) and transwell assays. In addition, the effects of TGIF2/FCMR axis on CCa metastasis were further explored in nude mice in vivo. Our data revealed a significant increase in TGIF2 mRNA expression in CCa tissue specimens compared to adjacent ones, and the increasing degree was positively correlated with the incidence of lymph node or distant metastasis of CCa patients. The results of CCK-8 and transwell suggested that knocking down TGIF2 effectively attenuated the proliferative ability and invasiveness of CCa cells. Luciferase assay confirmed that TGIF2 can directly bind to the DNA promoter of its target gene FCMR. Simultaneous transfection of sh-TGIF2 and sh-FCMR partially reversed the inhibitory effect of single transfection of TGIF2 knockdown on the malignant progression of CCa. Experiments in nude mice also suggested that TGIF2 could promote CCa tumorigenesis through the modulation of FCMR expression. In summary, TGIF2 can promote the migration and proliferation ability of cervical cancer cells via down-regulating FCMR. Our study provides a new therapeutic target for the clinical treatment of cervical cancer.

CFTR Regulates the Proliferation, Migration and Invasion of Cervical Cancer Cells by Inhibiting the NF-κB Signalling Pathway.

BackgroundThe cystic fibrosis transmembrane conductance regulator (CFTR) and nuclear factor κB (NF-κB) signalling pathways are currently regarded as co-regulators of the occurrence of cervical cancer. However, the detailed mechanism of CFTR- and NF-κB-mediated effects in cervical cancer remains to be elucidated. This study aimed to investigate the mechanism by which CFTR and NF-κB influence the development of cervical cancer.Patients and MethodsCFTR ΔF508 mutation and CFTR promoter methylation were detected in cervical tissue samples. NF-κB p65 and IκBα protein levels were tested in HeLa cells with CFTR overexpression and knockdown by Western blotting. The effects of CFTR on cell proliferation, migration, and invasion were examined in HeLa cells by WST-1 and soft agar assays, cell wound scratch assay, and Matrigel invasion assays, respectively. The protein–protein interaction (PPI) network was constructed by using GeneMANIA. GeneCoDis3 was used to perform Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) functional enrichment analysis on genes in the PPI network.ResultsCFTR mutation and CFTR promoter methylation were not associated with the occurrence of cervical cancer. NF-κB p65 protein levels were decreased in CFTR overexpression lines and increased in CFTR knockdown lines, and IκBα levels were affected in the opposite manner, indicating that CFTR inhibited the NF-κB signalling pathway. CFTR also regulated the cell proliferation, migration, and invasion ability of cervical cancer cells. When CFTR was overexpressed, cell proliferation, migration, and invasion ability were decreased. There were 20 genes that interacted with CFTR. KEGG pathway analysis showed enrichment in the gastric acid secretion, chemokine signalling, bile secretion and apoptosis pathways.ConclusionCFTR plays an important role in cancer cell proliferation, migration and invasion by inhibiting the NF-κB signalling pathway in cervical cancer.

Neferine, an alkaloid from lotus seed embryo targets HeLa and SiHa cervical cancer cells via pro-oxidant anticancer mechanism.

Apoptosis and autophagy are important processes that control cellular homeostasis and have been highlighted as promising targets for novel anticancer drugs. This study aims to investigate the inhibitory effects and mechanisms of Neferine (Nef), an alkaloid from the lotus seed embryos of Nelumbo nucifera (N. nucifera), as a dual inducer of apoptosis and autophagy through the reactive oxygen species (ROS) activation in cervical cancer cells. Nef and N. nucifera extract suppressed the cell viability of HeLa and SiHa cells in a dose-dependent manner. Importantly, Nef showed minimal toxicity to normal cells. Furthermore, Nef inhibited anchorage-independent growth, colony formation and migration ability of cervical cancer cells. Nef induces mitochondrial apoptosis by increasing pro-apoptotic protein bax, cytochrome-c, cleaved caspase-3 and caspase-9, poly-ADP ribose polymerase (PARP) cleavage, DNA damage (pH2 AX) while downregulating Bcl-2, procaspase-3 and procaspase-9, and TCTP. Of note, apoptotic effect by Nef was significantly attenuated in the presence of N-acetylcysteine (NAC), suggesting pro-oxidant activity of this compound. Nef also promoted autophagy induction through increasing beclin-1, atg-4, atg-5 and atg-12, LC-3 activation, and P 62/SQSTM1 as determined by western blot analysis. Collectively, these results demonstrate that Nef is a potent anticancer compound against cervical cancer cells through inducing apoptosis and autophagic pathway involving ROS.

A novel function of IMPA2, plays a tumor-promoting role in cervical cancer

Discovery of genes and molecular mechanism involved in cervical cancer development would promote the prevention and treatment. By comparing gene expression profiles of cervical carcinoma in situ (CCIS) and adjacent normal tissues, we identified a potential cancer-promoting gene, IMPA2. This study aimed to elucidate the role of IMPA2 and underlying molecular mechanisms in cervical cancer progression. To do this expression of IMPA2 was compared between human cervical cancer and corresponding adjacent normal cervical tissues firstly. CCK-8 assay, clone formation assay, wound healing assay, transwell assay, and tumor formation in nude mice were performed to demonstrate the effect of IMPA2 in cervical cancer proliferation and metastasis. Further proteomic profiling and western blotting explored the molecular pathway involved in the IMPA2-regulating process. The results showed that IMPA2 gene expression was upregulated in cervical cancer. Consistently, silencing of IMPA2 suppressed tumor formation in BALB/c nude mice. Short hairpin RNA (shRNA)-mediated IMPA2 silencing significantly inhibited proliferation and colony-forming abilities of cervical cancer cells, while IMPA2 overexpression had little impact. Also, IMPA2 silencing suppressed cellular migration, but overexpression promoted migration. Proteomics analysis revealed the involvement of mitogen-activated protein kinase (MAPK) pathway in tumor-promoting action of IMPA2. Significantly, the inhibition of IMPA2 activated ERK phosphorylation, and its inhibitory effects can be restored by using selective ERK inhibitor, FR180204. In conclusion, IMPA2 acts as an oncogene in the proliferation and migration of cervical cancer. IMPA2 downregulated ERK phosphorylation to promote cervical cancer. These findings identify a new mechanism underlying cervical cancer and suggest a regulating effect of IMPA2 in MAPK signaling pathway.

MiR‑96 exerts an oncogenic role in the progression of cervical cancer by targeting CAV‑1.

Cervical cancer is the 4th most common malignant tumor type affecting women worldwide; however, its molecular mechanisms are not fully understood. Previous studies have indicated that microRNAs (miRs) serve crucial roles in the cellular functions of tumors. miR‑96 is involved in the tumorigenesis of many cancer types. The aim of the present study was to investigate the role and mechanism of miR‑96 in the progression of cervical cancer. The present results suggested that overexpression of miR‑96 significantly enhanced the proliferative, migratory and invasive abilities of cervical cancer cells, while inhibiting miR‑96 had the opposite effects. Additionally, activation of the Akt/mTOR signaling pathway was enhanced by miR‑96 overexpression, while it was inhibited by the miR‑96 inhibitor. Moreover, it was identified that miR‑96 may directly target caveolin‑1 (CAV‑1) to decrease its expression level. Furthermore, overexpression of CAV‑1 could reverse the increase in cell proliferation, migration and invasion induced by miR‑96, as well as the upregulation of the Akt/mTOR signaling pathway. In conclusion, the present results suggested that miR‑96 may have an oncogenic role in the progression of cervical cancer by targeting CAV‑1. Therefore, miR‑96 may be a potential target for cervical cancer therapy.

CircAGFG1 aggravates the progression of cervical cancer by downregulating p53.

To explore the role of circAGFG1 in influencing the progression of cervical cancer (CC) and the underlying molecular mechanism. CircAGFG1 levels in CC tissues and paracancerous tissues were determined by quantitative Real Time-Polymerase Chain Reaction (qRT-PCR). Its level in CC cell lines was detected as well. Meanwhile, circAGFG1 levels in CC patients with different tumor staging, metastatic statues, and tumor sizes were examined. The Kaplan-Meier method was introduced for assessing the prognostic potential of circAGFG1 in CC. The regulatory effects of circAGFG1 on the proliferative ability of CC cells were evaluated by performing the Cell Counting Kit-8 (CCK-8) and 5-Ethynyl-2'-deoxyuridine (EdU) assay. The subcellular distribution of circAGFG1 in the CC cells was analyzed. Through chromatin immunoprecipitation (ChIP) and RNA immunoprecipitation (RIP) assay, the interaction between circAGFG1 and p53 was determined. Finally, the role of the circAGFG1/p53 axis in influencing the proliferation of CC cells was uncovered. CircAGFG1 was upregulated in CC tissues and cell lines. Besides, the circAGFG1 level was closely related to worse tumor staging, a higher rate of metastasis, and larger tumor size in CC patients. Besides, CC patients with a high level of circAGFG1 presented worse prognosis. The knockdown of circAGFG1 attenuated the proliferative ability of SiHa and HeLa cells. CircAGFG1 was mainly distributed in the nucleus of the CC cells. The interaction between circAGFG1 and p53 was verified. The knockdown of p53 could partially reverse the regulatory effect of circAGFG1 on the proliferative ability of the CC cells. CircAGFG1 is upregulated in CC. By recruiting EZH2, circAGFG1 downregulates p53 and thus exerts a carcinogenic role to accelerate the malignant progression of cervical cancer.

Ganoderma Lucidum Polysaccharide, an Extract from Ganoderma Lucidum, Exerts Suppressive Effect on Cervical Cancer Cell Malignancy through Mitigating Epithelial-Mesenchymal and JAK/STAT5 Signaling Pathway

Background/Aims: We aimed to explore whether ganoderma lucidum polysaccharide (GLP) exhibits antitumor effect on cervical cancer cells. Methods and Results: Different concentration of GLP was used to treat cervical cell. The data from cell counting kit-8 assay proved that the optimal working concentration and time of GLP were 200 µg/mL and treated for 48 h. The transwell assay demonstrated that GLP could attenuate the invasion and migration abilities of cervical cancer cells. Moreover, flow cytometry illustrated that GLP could promote the apoptosis of cervical cancer cells and limit the cycle of cervical cancer cells. Western blot assay discovered that the expression of proapoptosis proteins including Bax, Cleaved Caspases 3 and 9 increased and the antiapoptosis protein Bcl-2 decreased after treated with GLP. Moreover, we found that the expression of E-cadherin was increased, and N-cadherin, Vimentin, and Slug were decreased. Meanwhile, the expression of phosphorylated-JAK and phosphorylated-STAT5 was also decreased in cervical cancer cells treated by GLP, suggesting the inhibitory effect on JAK/STAT5 pathways. Conclusions: All of these data illustrated that GLP could alleviate the activity and aggressiveness, block the cell cycle, and promote the apoptosis of cervical cancer cells, which were possible via inhibiting epithelial-mesenchymal and JAK/STAT5 pathways.

Circular RNA hsa_circ_0007364 increases cervical cancer progression through activating methionine adenosyltransferase II alpha (MAT2A) expression by restraining microRNA-101-5p

ABSTRACT Emerging evidence suggested that circular RNAs (circRNAs) play critical roles in cervical cancer (CC) progression. However, the roles and molecular mechanisms of hsa_circ_0007364 in the tumorigenesis of CC remain unclear. In the present study, we used bioinformatics analysis and a series of experimental analysis to characterize a novel circRNA, hsa_circ_0007364 was up-regulated and associated with advanced clinical features in CC patients. Hsa_circ_0007364 inhibition notably suppressed the proliferation and invasion abilities of CC cells in vitro and reduced tumor growth in vivo. Moreover, hsa_circ_0007364 was uncovered to sponge miR-101-5p. Additionally, methionine adenosyltransferase II alpha (MAT2A) was verified as a target gene of miR-101-5p, and its downregulation reversed the inhibitory effects of hsa_circ_0007364 knockdown on CC progression. Therefore, we suggested that hsa_circ_0007364 might serve as an oncogenic circRNA in CC progression by regulating the miR-101-5p/MAT2A axis, which provides a potential therapeutic target to the treatment. Research highlights hsa_circ_0007364 was upregulated in CC hsa_circ_0007364 promoted CC cell progression hsa_circ_0007364/miR-101-5p/MAT2A axis in CC

LZAP promotes the proliferation and invasiveness of cervical carcinoma cells by targeting AKT and EMT.

Objective: To explore the relationship and mechanism of LZAP in the occurrence and development of cervical cancer and to provide a new target and intervention method for the treatment of cervical cancer.Methods: Data mining and analysis of LZAP expression levels were performed using several online databases, including The Cancer Genome Atlas (TCGA). A cervical cancer cell line that stably overexpresses LZAP was established, and the effect of LZAP overexpression on cell proliferation, invasion, migration and tumor formation in vivo as well as its mechanism were explored.Results: Our study shows that the expression of LZAP is upregulated in cervical cancer. The overexpression of LZAP can significantly promote the proliferation, colony formation, and invasion and migration abilities of cervical cancer cells. The tumorigenesis test in nude mice showed that overexpression of LZAP could promote the tumorigenicity of cervical cancer cells in vivo. LZAP could also promote the phosphorylation of AKT at position 473 and the epithelial-mesenchymal transition (EMT).Conclusion: The expression of LAZP is increased in cervical cancer, which can enhance the invasion, metastasis, and EMT in cervical cancer cells by promoting AKT phosphorylation.

Metformin Inhibit Cervical Cancer Migration by Suppressing the FAK/Akt Signaling Pathway.

Background:Metformin, an antidiabetic drug, has been previously reported to have anti-cancer activities. However, its role in the control of cancer cell migration remains elusive. Methods:To examine the possible effect of metformin on migration of cervical cancer cells. The related mechanisms were further determined by immunocytochemistry and Western’s blotting assay. Results:The results showed that metformin treatment substantially inhibited the migration ability of cervical cancer cells. Consistently, the filopodia and lamellipodia formation were depleted after exposure to metformin. The suppression of migration mediated through the regulatory proteins such as focal adhesion kinase (FAK), ATP-dependent tyrosine kinase (Akt), Rac1 and RhoA after metformin treatment. Conclusion:Metformin displays antimigration effects in cervical cancer cells by inhibiting filopodia and lamellipodia formation through the suppression of FAK, Akt and its downstream Rac1 and RhoA protein. We propose that metformin could be a novel potential candidate as an antimetastatic cancer drug in the cervical cancer management.

Abstract A055: Knockdown of E-cadherin gene in cervical cancer cells: An in vitro model for cancer stem cell phenotype?

Abstract Background: Cervical cancer is one of the leading causes of cancer related morbidity and mortality amongst women, especially in developing countries. Recent studies suggest that Cancer stem cells (CSC) are critically involved in regulating the chemo resistant behavior of cancer cells and a relapse of the disease. So targeting CSC-like cells along with the bulk tumor cells can be a promising approach for therapeutics. In this context, we demonstrated that knockdown of E-cadherin gene in cervical cancer cells can generate stem cell-like phenotype and induce epithelial to mesenchymal transition. Interestingly, knockdown of E-cadherin gene in normal keratinocytes did not induce CSC-like cells. Methods: In our study the CSC phenotype cells were isolated by side population (SP) assay from cervical cancer cells and immortalized keratinocytes. The expression of stem cell marker and EMT genes were confirmed by qRT-PCR and immunostainings. Further, we performed SiRNA-mediated knockdown of E-Cadherin (E-Cad) and studied the tumorsphere formation, metastasis and invasion ability of cervical cancer cells. Next, using RTPCR based gene expression analysis and tumorsphere formation we studied the stem–like properties of cancer cells. We also performed dose dependent cell viability assay using clinically relevant drugs and inhibitors to analyze the drug resistance behavior of the tumorspheres (cervical organoids). Results: We found that E-Cad knockdown induces EMT in cervical cancer cells with stem cell-like characteristics along with enhanced tumorsphere formation, metastasis and invasion ability. These cells not only possess CSC-like phenotype at transcriptional and translational level but also functionally as shown by drug resistance behavior studies. Conclusion: Thus, we believe that this can be a promising model to study CSC-like characteristics in-vitro in cervical cancer to target the molecular pathways for therapeutics. Citation Format: Shalmoli Bhattacharyya, Anuka Sharma, Arnab Pal, Radhika Srinivasan. Knockdown of E-cadherin gene in cervical cancer cells: An in vitro model for cancer stem cell phenotype? [abstract]. In: Proceedings of the AACR-NCI-EORTC International Conference on Molecular Targets and Cancer Therapeutics; 2019 Oct 26-30; Boston, MA. Philadelphia (PA): AACR; Mol Cancer Ther 2019;18(12 Suppl):Abstract nr A055. doi:10.1158/1535-7163.TARG-19-A055

CircAGFG1 promotes cervical cancer progression via miR-370-3p/RAF1 signaling

BackgroundIn past decades, circular RNAs (circRNAs) have achieved increasing attention because of its regulatory role in different kinds of cancers. However, how circAGFG1 regulates cervical cancer (CC) is still largely undiscovered. This study aims to evaluate the role of a novel circRNAs and related molecular mechanism in CC cells.MethodsHigh or low level of circAGFG1 was detected in CC cells or normal cell line with qRT-PCR. The proliferative and migratory abilities of CC cells were assessed with loss-of function assays. The downstream miRNA and mRNA of circAGFG1 were searched out and proved by using bioinformatics analysis and mechanism experiments. Recue assays were designed to confirm the role of circAGFG1/miR-370-3p/RAF1 axis in CC cell activities.ResultsThe levels of circAGFG1 was abundant in CC cells in comparison with normal cervical cell End1/E6E7. The inhibitory effect of decreased circAGFG1 level on the proliferative and migratory abilities of CC cells was assessed. CircAGFG1 and miR-370-3p were localized in the cytoplasm and they can interact with each other. Moreover, miR-370-3p was downregulated in CC cells. We also determined the negative effect of miR-370-3p on RAF1. CircAGFG1 could promote RAF1 expression by absorbing miR-370-3p, thereby activating RAF/MEK/ERK pathway. circAGFG1 promoted proliferation and migration of CC cells via enhancing the activity of RAF/MEK/ERK pathway by sponging miR-370-3p and further regulating RAF1.ConclusionThe results of this study provided new evidence that circAGFG1 acted as a vital regulator in cervical cancer proliferation and migration, giving great promise to apply it as a potential biomarker for diagnosis and therapy in CC treatment.

LINC00116 enhances cervical cancer tumorigenesis through miR-106a/c-Jun pathway.

Some studies imply that LINC00116 is involved in cervical cancer progression; however, the molecular mechanism by which LINC00116 modulating tumorigenesis of cervical cancer remains not clear. Reverse transcription-quantitative PCR (RT-qPCR)and the Western blot approaches were employed to probe genes expression levels. To examine the tumorigenic abilities of cervical cancer cells, MTT assay, Transwell assay, and wound-healing assay were used to investigate proliferation, invasion, and migration of HeLa or C-33A cells. LINC00116 knockdown attenuates cell proliferation, invasion, and migration of cervical cancer cells. miR-106a directly binds LINC00116 and regulate each other. Moreover, miR-106a inhibitor remarkably enhanced tumorigenesis of shLINC00116 HeLa cells. Through bioinformatic and dual-luciferase reporter assay, the results showed that miR-106a mimic directly targeted and downregulated the c-Jun. c-Jun overexpression could greatly rescue miR-106a mimic-modulated cervical cancer tumorigenesis. LINC00116 knockdown and miR-106a mimic-modulated programmed cell death ligand 1 (PD-L1) expression, which could be reverted by c-Jun introduction. LINC00116, PD-L1, and JUN were both upregulated in cervical cancer tumors compared to normal tissues. Lower expression levels of LINC00116 and JUN, as well as higher level of miR-106a were closely associated with higher overall survival of cervical cancer patients. Here, we report a novel role for LINC00116 in tumorigenesis of cervical cancer by regulating miR-106a/c-Jun axis. Our findings provide a foundation for understanding cervical cancer and facilitate the development of therapeutical approaches by targeting LINC00116.

LncRNATCF7 up-regulates DNMT1 mediated by HPV-18 E6 and regulates biological behavior of cervical cancer cells by inhibiting miR-155.

This work aimed to study the mechanism of lncRNATCF7 upregulating DNMT1 mediated by HPV-18 E6 and regulating the biological behavior of cervical cancer cells by inhibiting miR-155. HPV-16 E6 enhanced DNMT1 expression in cervical cancer cells, which was detected by Western blotting. The expression of miR-155 in cervical cancer was detected by qPCR, the interaction between TCF-7 and miR-155 by Dual-luciferase reporter gene. The changes in invasion ability of cervical cancer cells and the effect of miR-155 on the invasion ability of cervical cancer cells after inhibiting TCF-7 were detected by the transwell invasion assay, while changes in migration ability of cervical cancer cells and the effect of miR-155 on migration ability of cervical cancer cells after inhibiting TCF-7 were observed by the scratch assay. The effect of inhibiting TCF-7 on the tumor size and volume of cervical cancer was detected by the subcutaneous tumor formation in nude mice. E6 expression was significantly inhibited by E6 siRNA. The knockdown of endogenous HPV-16 E6 markedly inhibited the expression of DNMT1; TCF-7 specifically bound to the 3' UTR of miR-155; inhibition of TCF-7 can inhibit invasion and migration of cervical cancer cells; enhanced miR-155 after the inhibition of TCF-7 can promote the invasion and migration of cervical cancer cells; compared with NC group, the tumor volume and weight of TCF-7-siRNA group tumor-bearing was significantly reduced. TCF-7 plays an important role in the development of cervical cancer. TCF-7 can target miR-155 to regulate the invasion and migration of cervical cancer cells.

Circular RNA circRNA_0000285 promotes cervical cancer development by regulating FUS.

Recently, the vital role of circular RNAs (circRNAs) in human diseases has attracted much attention. The aim of this research was to verify the potential role of circRNA_0000285 in the development of cervical cancer (CC). CircRNA_0000285 expression in both CC cells and tissue samples was detected by quantitative Real Time-Polymerase Chain Reaction (qRT-PCR). Functional experiments were performed, including cell counting kit-8 (CCK-8) assay, cell cycle assay and transwell assay. Meanwhile, the underlying mechanism was explored through qRT-PCR and Western blot assay, respectively. In addition, the function of circRNA_0000285 was identified in vivo. CircRNA_0000285 expression level was significantly higher in CC samples than that of corresponding normal tissues. Moreover, the growth and migration abilities of CC cells were significantly inhibited after circRNA_0000285 was knocked down in vitro. Furthermore, the expression of FUS was remarkably down-regulated after knockdown of circRNA_0000285. Subsequent results indicated that the expression level of FUS was positively correlated with the expression of circRNA_0000285 in CC tissues. In addition, the knockdown of circRNA_0000285 significantly inhibited the formation and metastasis of CC in nude mice. CircRNA_0000285 could enhance the proliferation and metastasis of CC by up-regulating FUS, which might be a potential therapeutic target for CC treatment.

Knockdown of long noncoding RNA DLX6-AS1 inhibits cell proliferation and invasion of cervical cancer cells by downregulating FUS.

Recently, the vital role of long noncoding RNAs (lncRNAs) in human diseases have got much attention. In this research, lncRNA DLX6-AS1 is studied to verify how it affects the development of cervical cancer (CC). DLX6-AS1 expression was detected by quantitative Real Time-Polymerase Chain Reaction (qRT-PCR) in both CC cells and tissue samples. Besides, functional experiments including cell counting kit-8 (CCK-8) assay, colony formation assay, and transwell assay were performed. Meanwhile, the underlying mechanism was explored through qRT-PCR and Western blot assay. The function of DLX6-AS1 was also identified in vivo. By comparing with corresponding tissues, the DLX6-AS1 expression level was significantly higher in CC samples. Moreover, cell growth ability and invaded ability of CC cells were inhibited after DLX6-AS1 was knocked down. Furthermore, the expression of FUS was inhibited after knockdown of DLX6-AS1. It was found that the expression level of FUS positively correlated to the expression of DLX6-AS1 in CC tissues. In addition, knockdown of DLX6-AS1 inhibited tumor formation and metastasis of CC in nude mice. These results suggest that DLX6-AS1 could enhance cell proliferation and metastasis of CC by upregulating FUS, which might be a potential therapeutic target in CC.

ELK1-activated GPC3-AS1/GPC3 axis promotes the proliferation and migration of cervical cancer cells.

Long non-coding RNA (lncRNA) is reported to be involved in multiple biological processes in numerous human tumors. Furthermore, an increasing number of studies have confirmed the involvement of lncRNA in the initiation and development of human cancers, including cervical cancer (CC). The present study explored the potential role of lncRNA glypican 3 antisense transcript 1 (GPC3-AS1) with respect to regulating CC cell proliferation and migration. A quantitative reverse transcriptase-polymerase chain reaction confirmed that GPC3-AS1 was up-regulated in CC cells compared to normal CRL-2614 cells. Loss-of-function assays demonstrated the negative effect of GPC3-AS1 depletion on CC cell proliferation and migration. GPC3-AS1 positively regulated its nearby gene glypican 3 (GPC3). Significant up-regulation of GPC3 was also observed in CC cells, consistently with GPC3-AS1. In addition, GPC3-AS1 and GPC3 synergistically promoted the proliferative and migratory abilities of CC cells. Mechanistic investigation showed that ELK1 acted as the transcription activator of GPC3-AS1 and GPC3, thus contributing to their up-regulation in CC cells. Rescue assays indicated that the ELK1-induced GPC3-AS1/GPC3 axis promoted cell proliferation and migration in CC. The results of the present study have revealed a novel molecular pathway that can regulate CC cell proliferation and migration, thus providing a new basis for investigating the molecular mechanism associated with CC progression.

Long non-coding RNA MEG3 inhibits cervical cancer cell growth by promoting degradation of P-STAT3 protein via ubiquitination

BackgroundMaternally expressed 3 (MEG3) plays an important role in cervical cancer development, but its exact role remains unclear. Here, we explored the specific regulatory mechanism of MEG3 and its downstream proteins in cervical cancer cells.MethodsThe effect of MEG3 on tumor formation ability of cervical cancer cells was determined in nude mice. The direct binding of MEG3 to phosphorylated signal transducer and activator of transcription 3 (P-STAT3) was detected by RNA pull-down and RNA-binding protein immunoprecipitation (RIP) assays. Cycloheximide (CHX)-chase and ubiquitination assays were performed to determine the regulatory effect of MEG3 on P-STAT3 ubiquitination. Clone formation assay and flow cytometry were used to evaluate the effect of the MEG3-STAT3 regulatory axis on cell proliferation and apoptosis.ResultsIn vivo tumor formation experiments showed that MEG3 inhibited the tumor formation ability of cervical cancer cells. RNA pull-down and RIP assays demonstrated that MEG3 bound directly to P-STAT3 protein. CHX-chase and ubiquitination assay results showed that MEG3 promoted P-STAT3 degradation via ubiquitination. Clone formation assay and flow cytometry analysis results revealed that the inhibitory effect of MEG3 on P-STAT3 promoted apoptosis and inhibited proliferation of cervical cancer cells.ConclusionMEG3 binds to P-STAT3 in cervical cancer cells, resulting in P-STAT3 ubiquitination and degradation and apoptosis and inhibition of proliferation of tumor cells. The in-depth elaboration of the MEG3-STAT3 regulatory axis in cervical cancer may clarify the mechanism of action of MEG3 and provide new ideas for cervical cancer treatment.

LncRNA TP73-AS1 is a novel regulator in cervical cancer via miR-329-3p/ARF1 axis.

Cervical cancer holds one of the highest morbidity and mortality in various types of cancers. It even leads to the most number of cancer-related deaths of women. A lot of research has indicated that the anomalous expression of long noncoding RNAs (lncRNAs) would induce carcinogenesis and is associated with poor prognosis of patients with cancer. However, the function and mechanism of many lncRNAs still call for further research. Tumor Protein P73 Antisense RNA 1 (TP73-AS1) is no exception. LncRNA TP73-AS1 has been found to promote cancer progressions in various cancers. It is upregulated in cervical cancer cells. The proliferation and migration ability of cervical cancer cells can also be boosted by TP73-AS1 in return. Meanwhile, miRNA-329-3p is downregulated in cervical cancer cells and could bind with both TP73-AS1 and ADP Ribosylation Factor 1 (ARF1). TP73-AS1 inhibited miR-329-3p expression while miR-329-3p inhibited ARF1 expression. More importantly, TP73-AS1 can positively regulate ARF1 expression. Based on all these experiments, TP73-AS1 regulates ARF1 expression by competitively binding with miR-329-3p, thus regulating cervical cancer progression. Further rescue assays confirmed TP73-AS1 regulates cervical cell proliferation and migration via miR-329-3p/ARF1. TP73-AS1 might serve as a novel regulator in cervical cancer.

CircMTO1 promotes tumorigenesis and chemoresistance of cervical cancer via regulating miR-6893

BackgroundCervical cancer has been one of the most common cancer in women worldwide. However, the detailed mechanism underlying circMTO1-regulated cervical cancer remains unclear. MethodsRT-qPCR and westernblot were used to examine genes expression levels. Wound healing assay and transwell invasion assay were utilized to probe migration and invasion abilities of cervical cancer cells. In addition, cell viability and apoptosis were measured by MTT and TUNEL assay, respectively. Finally, we employed bioinformatic approach to analyze gene expression levels in clinical cervical cancer tissues. ResultscircMTO1 was shown to be greatly upregulated in cervical cancer cell lines and tumors. circMTO1 directly interacts with miR-6893. miR-6893 could restore circMTO1-regulated migration, invasion and chemoresistance of cervical cancer cells. Furthermore, we identified S100A1 as downstream effector for circMTO1/miR-6893-induced tumorigenesis of cervical cancer cells. Besides, westernblot analyses demonstrated that circMTO1 and miR-6893 inhibitor enhanced Beclin1 expression and downregulated p62 level. We found that autophagy inhibitor 3-MA could revert cell viability and apoptosis of HeLa regulated by circMTO1 and miR-6893 inhibitor. More importantly, the bioinformatic results showed the dysregulated expression patterns of circMTO1, miR-6893 and S100A1 in human clinical cervical cancer tissues. ConclusionsIn summary, we reveal, for the first time, a novel mechanism of circMTO1/miR-6893 in tumorigenesis and chemoresistance of cervical cancer. Our findings support the notion of developing the new therapies and biomarkers by targeting circMTO1 for cervical cancer.