scholarly journals Effect of miRNA-200b on the proliferation and apoptosis of cervical cancer cells by targeting RhoA

Open Medicine ◽  
2020 ◽  
Vol 15 (1) ◽  
pp. 1019-1027
Author(s):  
Lijie He ◽  
Jing Wang ◽  
Dandan Chang ◽  
Dandan Lv ◽  
Haina Li ◽  
...  
Keyword(s):  
Cervical Cancer ◽  
Cancer Cells ◽  
Negative Control Group ◽  
Negative Control ◽  
Luciferase Reporter ◽  
Control Group ◽  
Rt Pcr ◽  
Blank Control Group ◽  
Cervical Cancer Cells ◽  
Mrna And Protein Expression

AbstractObjectiveThis article aims to investigate the effect of miRNA-200b on the proliferation and apoptosis of cervical cancer cells by targeting RhoA.MethodsHeLa cells of cervical cancer were divided into five groups: blank control group, negative control group (miRNA-200b mimic NC), miRNA-200b mimic group, RhoA-negative control group, and RhoA overexpression group. Cells were collected 48 h after transfection. The expression levels of miRNA-200b were detected by RT-PCR. Target relationship between miRNA-200b and RhoA was verified by the dual-luciferase reporter assay. RhoA mRNA and protein expression were detected by western blot and RT-PCR methods. Flow cytometry was used to detect the apoptosis of cells in each group, and the CCK8 method was used to detect the proliferation of cells in each group. The mRNA and protein expression of Bax and cyclin D1 were detected by RT-PCR and western blot.ResultsThe results of the dual luciferase reporter assay showed that RhoA was the target gene of microRNA 200b. Compared with the blank control group and the miRNA-200b mimic-NC group, the proportion of apoptotic cells increased significantly in the miRNA-200b mimic group, and the proliferation of cells was inhibited (P < 0.05). After overexpression of RhoA, the percentage of apoptotic cells decreased and the ability of cell proliferation increased significantly (P < 0.05).ConclusionmiRNA-200b can inhibit the proliferation and promote the apoptosis of cervical cancer cells by targeting the RhoA gene.

scholarly journals MicroRNA-96-5p Facilitates the Viability, Migration, and Invasion and Suppresses the Apoptosis of Cervical Cancer Cells byNegatively Modulating SFRP4

2020 ◽  
Vol 19 ◽  
pp. 153303382093413 ◽  
Author(s):  
Huiling Zhang ◽  
Ruxin Chen ◽  
Jinyan Shao
Keyword(s):  
Cervical Cancer ◽  
Cancer Cells ◽  
Clinical Stage ◽  
Luciferase Reporter ◽  
Migration And Invasion ◽  
Related Protein ◽  
Siha Cells ◽  
Gene Assay ◽  
Cervical Cancer Cells ◽  
Cancer Tissues

Purpose: The current study was intended to research the functional role and regulatory mechanism of microRNA-96-5p in the progression of cervical cancer. Methods: MicroRNA-96-5p expression in cervical cancer tissues was assessed by quantitative real-time polymerase chain reaction. The association between microRNA-96-5p expression and clinicopathological features of patients with cervical cancer was analyzed. MTT, flow cytometry, wound healing, and transwell assay were performed to evaluate the viability, apoptosis, migration, and invasion of Hela and SiHa cells. Targetscan, dual-luciferase reporter gene assay, and RNA pull-down analysis were constructed to evaluate the target relationship between microRNA-96-5p and secreted frizzled-related protein 4. Results: MicroRNA-96-5p was overexpressed in cervical cancer tissues, and microRNA-96-5p expression was markedly associated with the clinical stage and lymph node metastasis of patients with cervical cancer. Overexpressed microRNA-96-5p facilitated the viability, migration, invasion, and inhibited the apoptosis of Hela and SiHa cells, whereas suppression of microRNA-96-5p exerted the opposite trend. Secreted frizzled-related protein 4 was proved to be a target of microRNA-96-5p. Silencing of secreted frizzled-related protein 4 eliminated the anti-tumor effect of microRNA-96-5p on cervical cancer cells. Conclusions: MicroRNA-96-5p facilitated the viability, migration, and invasion and inhibited the apoptosis of cervical cancer cells via negatively regulating secreted frizzled-related protein 4.

scholarly journals HAND2-AS1 inhibits invasion and metastasis of cervical cancer cells via microRNA-330-5p-mediated LDOC1

2019 ◽  
Vol 19 (1) ◽  
Author(s):  
Shengcai Chen ◽  
Jing Wang
Keyword(s):  
Cervical Cancer ◽  
Lymph Node ◽  
Lymph Node Metastasis ◽  
Cancer Cells ◽  
Cell Counting ◽  
Luciferase Reporter ◽  
Invasion And Metastasis ◽  
Node Metastasis ◽  
Cervical Cancer Cells ◽  
Associated Proteins

Abstract Background Cervical cancer is a serious disease with complicated pathogenesis and thus there is an urgent need to find novel targets for the treatment. Recently, long non-coding RNAs (lncRNAs) have emerged as critical factors in tumorigenesis. In this study, we aimed to investigate the mechanism of HAND2 antisense RNA 1 (HAND2-AS1) on the invasion and metastasis of cervical cancer cells. Methods The expression patterns of HAND2-AS1, microRNA-330-5p (miR-330-5p) and leucine zipper down-regulated in cancer 1 (LDOC1) in cervical cancer were characterized by RT-qPCR and western blot analysis. Dual luciferase reporter assay and RIP were applied to verify relationship between HAND2-AS1, miR-330-5p and LDOC1. Fluorescence in situ hybridization (FISH) was used to detect the subcellular localization of HAND2-AS1. Besides, viability, invasion and migration ability of HeLa cells were investigated by cell counting kit-8 (CCK-8) and Transwell assays respectively. Hematoxylin–eosin staining was performed for lymph node metastasis detection. In addition, the tumor growth in nude mice was evaluated. Results Low expression of HAND2-AS1 and LDOC1, and high expression of miR-330-5p were detected in cervical cancer tissues and cells. It was found that binding of HAND2-AS1 to miR-330-5p results in upregulation of LDOC1 expression. Also, overexpressed HAND2-AS1 and LDOC1 or down-regulated miR-330-5p inhibited expression of proliferation-associated proteins Ki-67, PCNA, migration-associated proteins N-cad and invasion-related proteins MMP-2, MMP-9 as well as lymph node metastasis. Moreover, HAND2-AS1 inhibited tumor formation and lymph node metastasis by binding to miR-330-5p in vivo. Conclusion HAND2-AS1 promotes LDOC1 expression by competitively binding to miR-330-5p and consequently inhibiting cervical cancer cell invasion and metastasis. This could facilitate development of therapeutic strategies against cervical cancer.

scholarly journals Cellular Levels of Oxidative Stress Affect the Response of Cervical Cancer Cells to Chemotherapeutic Agents

2014 ◽  
Vol 2014 ◽  
pp. 1-14 ◽  
Author(s):  
Maria Filippova ◽  
Valery Filippov ◽  
Vonetta M. Williams ◽  
Kangling Zhang ◽  
Anatolii Kokoza ◽  
...  
Keyword(s):  
Oxidative Stress ◽  
Cervical Cancer ◽  
Cancer Cells ◽  
Adaptive Systems ◽  
Chemotherapeutic Agents ◽  
Rt Pcr ◽  
Cellular Models ◽  
Siha Cells ◽  
Ros Metabolism ◽  
Cervical Cancer Cells

Treatment of advanced and relapsed cervical cancer is frequently ineffective, due in large part to chemoresistance. To examine the pathways responsible, we employed the cervical carcinoma-derived SiHa and CaSki cells as cellular models of resistance and sensitivity, respectively, to treatment with chemotherapeutic agents, doxorubicin, and cisplatin. We compared the proteomic profiles of SiHa and CaSki cells and identified pathways with the potential to contribute to the differential response. We then extended these findings by comparing the expression level of genes involved in reactive oxygen species (ROS) metabolism through the use of a RT-PCR array. The analyses demonstrated that the resistant SiHa cells expressed higher levels of antioxidant enzymes. Decreasing or increasing oxidative stress led to protection or sensitization, respectively, in both cell lines, supporting the idea that cellular levels of oxidative stress affect responsiveness to treatment. Interestingly, doxorubicin and cisplatin induced different profiles of ROS, and these differences appear to contribute to the sensitivity to treatment displayed by cervical cancer cells. Overall, our findings demonstrate that cervical cancer cells display variable profiles with respect to their redox-generating and -adaptive systems, and that these different profiles have the potential to contribute to their responses to treatments with chemotherapy.

Effect of Zanthoxylum acanthopodium methanol extract on CDK4 expression to cervical cancer

2021 ◽  
pp. 5647-5652
Author(s):  
Syafruddin Ilyas ◽  
Rostime H. Simanullang ◽  
Salomo Hutahaean ◽  
Rosidah Rosidah ◽  
Putri C. Situmorang
Keyword(s):  
Cervical Cancer ◽  
Cancer Cells ◽  
Methanol Extract ◽  
Cancer Drug ◽  
Drug Candidate ◽  
Control Group ◽  
Cervical Tissue ◽  
Cervical Cancer Cells ◽  
Clone Development ◽  
Cdk4 Expression

Cervical cancer is a disease from the Human papillomavirus (HPV) through transmission, virus persistence, clone development until infecting the cells in the cervical. This study is to determine CDK4 expression in cervical cancer cells after being given Zanthoxylum acanthopodium methanol extract (ZAM) and the histological description of cervical cancer cells. This study consisted of 5 treatment groups. K-: control group, K+: rats model of cancer, P1: rats model of cancer with a dose of 100mg/BW of ZAM, P2: rats model of cancer with a dose of 200 mg/BW of ZAM, and P3: rats model of cancer with a dose of 400 mg/BW of ZAM. The cervical tissue was prepared on paraffin blocks and given Immunohistochemistry staining. Results showed that the expression of CDK4 are reduced in the ZAM treatment at doses of 200 and 400mg/KgBW (P<0.05) in cervical histology, but in doses of 100mg/kg BW, many brown marks are still visible on the cervical tissue. These proteins will bind, inhibit proteins, cell cycle development, modulate cell division, and signal transduction pathways of apoptotic signaling. The injection of benzopyrene and given ZAM in cervical tissue affect hematological values. ZAM affects and improves cervical histology after benzopyrene injection. The extract of andaliman can be developed into a cervical cancer drug candidate.

scholarly journals Up-regulation of miRNA-148a inhibits proliferation, invasion, and migration while promoting apoptosis of cervical cancer cells by down-regulating RRS1

2019 ◽  
Vol 39 (5) ◽  
Author(s):  
Ying Zhang ◽  
Bingmei Sun ◽  
Lianbin Zhao ◽  
Zhengling Liu ◽  
Zonglan Xu ◽  
...  
Keyword(s):  
Cervical Cancer ◽  
Cancer Cells ◽  
Cell Line ◽  
Hela Cells ◽  
Luciferase Reporter ◽  
Migration And Invasion ◽  
Invasion And Migration ◽  
Cervical Cancer Cells ◽  
And Migration

Abstract The purpose of the present study is to figure out the role of miRNA-148a (miR-148a) in growth, apoptosis, invasion, and migration of cervical cancer cells by binding to regulator of ribosome synthesis 1 (RRS1). Cervical cancer and adjacent normal tissues, as well as cervical cancer cell line Caski, HeLa, C-33A, and normal cervical epithelial cell line H8 were obtained to detect the expression of miR-148a and RRS1. Relationship between miR-148a and RRS1 expression with clinicopathological characteristics was assessed. The selected Caski and HeLa cells were then transfected with miR-148a mimics, miR-148a inhibitors or RRS1 siRNA to investigate the role of miR-148a and RRS1 on proliferation, apoptosis, colony formation, invasion, and migration abilities of cervical cancer cells. Bioinformatics information and dual luciferase reporter gene assay was for used to detect the targetting relationship between miR-148a and RRS1. Down-regulated miR-148a and up-regulated RRS1 were found in cervical cancer tissues and cells. Down-regulated miR-148a and up-regulated RRS1 are closely related with prognostic factors of cervical cancer. RRS1 was determined as a target gene of miR-148a and miR-148a inhibited RRS1 expression in cervical cancer cells. Up-regulation of miR-148a inhibited cell proliferation, migration, and invasion while promoting apoptosis in Caski and HeLa cells. Our study suggests that miR-148a down-regulates RRS1 expression, thereby inhibiting the proliferation, migration, and invasion while promoting cell apoptosis of cervical cancer cells.

Epigenetic regulation of KLK7 gene expression in pancreatic and cervical cancer cells

2016 ◽  
Vol 397 (11) ◽  
pp. 1135-1146 ◽  
Author(s):  
Ilangovan Raju ◽  
Gur P. Kaushal ◽  
Randy S. Haun
Keyword(s):  
Cervical Cancer ◽  
Cancer Cells ◽  
Cancer Cell ◽  
Promoter Region ◽  
Luciferase Reporter ◽  
Mrna Levels ◽  
Reporter Activity ◽  
Promoter Deletion Analysis ◽  
Cervical Cancer Cells ◽  
Chip Analysis

Abstract Kallikrein-related peptidase 7 (KLK7) is a serine protease encoded within the kallikrein gene cluster located on human chromosome region 19q13.3-13.4. KLK7 is overexpressed in human pancreatic ductal adenocarcinomas (PDACs), but not in normal pancreas. Examination of KLK7 mRNA levels in pancreatic cancer cell lines revealed that it is readily detected in MIA PaCa-2 and PK-1 cells, but not in Panc-1 cells. Treatment of Panc-1 cells with the histone deacetylase (HDAC) inhibitor trichostatin A (TSA) significantly induced KLK7 mRNA expression. Similarly, KLK7 is highly expressed in cervical cancer cells, but its expression in the human cervical cancer cell line HeLa is only detected following TSA treatment. Promoter deletion analysis revealed that the proximal -238 promoter region, containing a putative Sp1-binding site, was sufficient for TSA activation of luciferase reporter activity, which was abrogated by the disruption of the Sp1-binding sequence. Consistent with the notion that TSA induced KLK7 expression via Sp1, co-expression of Sp1 with the KLK7-promoter/luciferase construct produced a significant increase in reporter activity. Chromatin immunoprecipitation (ChIP) analysis revealed enriched Sp1 occupancy on the KLK7 promoter following TSA treatment. Similarly, ChIP analysis showed the histone active mark, H3K4Me3, in the KLK7 promoter region was significantly increased after exposure to TSA.

scholarly journals MicroRNA-373-3p inhibits the growth of cervical cancer by targeting AKT1 both in vitro and in vivo

2021 ◽  
Author(s):  
Min-Min Yu ◽  
Gen-ju Wang ◽  
Kai-Hua Wu ◽  
Song-Lin Xue ◽  
Li- Li Ju ◽  
...  
Keyword(s):  
Cervical Cancer ◽  
Cancer Cells ◽  
Tumor Model ◽  
Cell Counting ◽  
Over Expression ◽  
Cervical Carcinoma Cell Line ◽  
Cervical Cancer Cells ◽  
Mrna And Protein Expression

Objective: In this study, we aimed to investigate the function of microRNA-373-3p (miR-373-3p) in the pathogenesis of cervical cancer. Methods: Human and mouse cervical cancer cell lines were transfected with miR-373-3p mimic and inhibitor. Cell proliferation and viability were evaluated with Cell Counting Kit-8 (CCK-8) assay and Lactate Dehydrogenase (LDH) assay, respectively. The AKT1-targeting role of miR-373-3p was analyzed by qPCR and Western blot. Finally, a mouse xenograft cervical tumor model was adopted to study the in vivo effect of miR-373-3p on tumor growth and the expression of AKT1. Results: Over-expression of miR-373-3p significantly reduced the proliferation of cervical carcinoma cell line in vitro. In addition, miR-373-3p overexpression also inhibited cervical cancer growth in tumor-bearing mice. Mechanistically, we found that AKT1 gene can be targeted by miR-373-3p. MiR-373-3p mimic decreased the mRNA and protein expression of AKT1, while the miR-373-3p inhibitor increased the level of AKT1 in cervical cancer cells. AKT1 overexpression rescued the proliferation of cervical cancer cells transfected with miR-373-3p. Conclusion: MiR-373-3p can serve as a novel anti-tumor microRNA in cervical cancer by targeting AKT1.

Inhibiting the Redox Function of APE1 Suppresses Cervical Cancer Metastasis via Disengagement of ZEB1 from E-cadherin in EMT

2021 ◽  
Author(s):  
qing li ◽  
Zhi-Wei Zhou ◽  
Wei Duan ◽  
Cheng-Yuan Qian ◽  
Shu-Nan Wang ◽  
...  
Keyword(s):  
Cervical Cancer ◽  
Lymph Node ◽  
Cancer Cells ◽  
Cancer Metastasis ◽  
Gene Set Enrichment Analysis ◽  
Negative Regulator ◽  
Luciferase Reporter ◽  
Dependent Manner ◽  
Cervical Cancer Cells ◽  
E Cadherin

Abstract Background: Metastasis is a major challenge in cervical cancer treatment. Previous studies have shown that the dual functional protein apurinic/apyrimidinic endonuclease 1 (APE1) promotes tumor metastasis and is overexpressed in cervical cancer. However, the biological role and mechanism of APE1 in cervical cancer metastasis have rarely been studied.Methods: We used gene set enrichment analysis (GSEA) to determine the APE1-related signaling pathways in cervical cancer. To investigate the role and mechanism of APE1 in cervical cancer metastasis, invasion, immunohistochemistry, immunofluorescence, western blotting, secondary structure prediction, coimmunoprecipitation, luciferase reporter, and electrophoretic mobility shift assays were performed. The inhibitory effects of the APE1 redox function inhibitor APX3330 on cervical cancer metastasis were evaluated using animal models.Results: Clinical data showed that high expression of APE1 was associated with lymph node metastasis in cervical cancer patients. GSEA results showed that APE1 was associated with epithelial to mesenchymal transition (EMT) in cervical cancer. Ectopic expression of APE1 promoted EMT and invasion of cervical cancer cells, whereas inhibition of APE1 suppressed EMT and invasion of cervical cancer cells in a redox function-dependent manner. Notably, APE1 redox function inhibitor APX3330 treatment dramatically suppressed cervical cancer cell lymph node and distant metastasis in vivo. Furthermore, we found that APE1 enhanced the interaction between ZEB1 and the E-cadherin promoter by binding to ZEB1, thereby suppressing the expression of E-cadherin, a negative regulator of EMT.Conclusion: Our findings help to elucidate the role played by APE1 in cervical cancer metastasis and targeting APE1 redox function may be a novel strategy for inhibiting cervical cancer metastasis.

Effects of Liposomal Nanoparticles-Mediated miR-126 on Cervical Cancer Cells via Anti-Programmed Cell Death-1/Programmed Death Ligand 1 (PD-1/PD-L1) Signaling

2021 ◽  
Vol 13 (8) ◽  
pp. 1506-1511
Author(s):  
Minjuan Xu ◽  
Jun Huang ◽  
Liefeng Wang
Keyword(s):  
Cervical Cancer ◽  
Cell Proliferation ◽  
Cancer Cells ◽  
Molecular Mechanisms ◽  
Control Group ◽  
Tumor Mass ◽  
Programmed Death ◽  
Cervical Cancer Cells ◽  
Signaling Inhibitors ◽  
Liposomal Nanoparticles

Cervical cancer is often treated with surgery, radiotherapy and chemotherapy, but it does not have the advantages of precise treatment and prognosis is not ideal. Molecular targeted therapy can make up for the above shortcomings. This study mainly analyzed the influence of miR-126 on cervical cancer cells and possible molecular mechanisms, so as to provide a reference for better clinical improvement of prognosis for cervical cancer. C33a cells were assigned into control group, empty carrier group (C33a cells were co-cultured with liposome nanoparticle carrier), inhibitor group (C33a cells were treated with PD-1/PD-L1 signaling pathway inhibitor), miR-126 group (miR-126 with liposomal nanoparticles as carrier was added to C33a cells), followed by expression analysis of miR-126 and AK2, cell proliferation, PD-1/PD-L1 signaling and phosphorylation levels, as well as tumor mass and volume in nude mice. At 24 h, no difference of cell proliferation was found (P > 0.05) but cell proliferation showed significant differences after cell growth of 48 h, with lower proliferation in inhibitor group and miR-126 group (P < 0.05). The levels of PD-1, PD-L1, AK2, and p-PD-1 in inhibitor group and miR-126 group were significantly lower than for the other two groups (P > 0.05). There was a target relationship between miR-126 and AK2. The transplanted tumor in the miR-126 group was significantly decreased, with lower tumor mass and volume than control group (P < 0.05). The carrier-based miR-126 and PD-1/PD-L1 signaling inhibitors can inhibit cervical cancer cell proliferation.

Sevoflurane Inhibits Growth and Metastasis of Cervical Cancer Through Up-Regulating miR-203 by Targeting Tumor Protein Translationally Controlled 1

2020 ◽  
Vol 10 (6) ◽  
pp. 874-883
Author(s):  
Li Zhang ◽  
Shiyou Wei ◽  
Zhenkai Xu ◽  
Wen Sun ◽  
Lihua Hang
Keyword(s):  
Cervical Cancer ◽  
Cell Viability ◽  
Cancer Cells ◽  
Target Genes ◽  
Luciferase Reporter ◽  
Migration And Invasion ◽  
Reporter System ◽  
Cervical Cancer Cells ◽  
Induced Apoptosis ◽  
Cancer Tissues

Background: Cervical cancer is a type of malignancy with high incidence and high mortality in women all over the world. Recent findings revealed the role of sevoflurane in the inhibition of development of various cancer types. This study aimed to explore whether sevoflurane could suppress cells proliferation and metastasis through adjusting miR-203 expression in cervical cancer. Methods: The effects of sevoflurane on HeLa cell viability was assessed using CCK-8 assay. miR-203 level in Hela cells was determined by qRT-PCR. In addition, cells apoptosis, migration and invasion were evaluated using flow cytometry and transwell analysis respectively after sevoflurane treatment or miR-203 expression changes. Bioinformatics software (TargetScan) was used to predict the potential target genes for miR-203 and the prediction was validated using dual-luciferase reporter system. Results: Sevoflurane effectively inhibited cell viability, metastasis and stimulated apoptosis in cervical cancer. miR-203 demonstrated a low expression in cervical cancer tissues and cells and sevoflurane significantly up-regulated miR-203 expression in cervical cancer cells. Upregulation of miR-203 significantly suppressed cell growth and metastasis and induced apoptosis, while down-regulation of miR-203 presented the opposite effects in cervical cancer cells. In addition, the inhibitory effects of sevoflurane were eliminated by down-regulating miR-203 in cervical cancer cells. In addition, TPT1 was confirmed as a target gene for miR-203. Conclusion: Sevoflurane inhibited cervical cancer cells viability and metastasis through up-regulation of miR-203 expression by targeting TPT1.

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