Knockdown of RNF6 Inhibits Cervical Cancer HeLa Cells Growth by Suppressing the MAPK/ERK Signaling

Abstract Background Given its crucial role in human malignancies, how Ring finger protein 6 (RNF6) functions in cervical cancer has yet to be elucidated. In our research, we explored the biological significance of RNF6 in cervical cancer HeLa cells and its possible regulatory mechanism. Methods The expression levels of RNF6 mRNA and protein in cervical cancer tissues and cells were both analyzed, the former by Gene Expression Profiling Interactive Analysis (GEPIA), and the latter by quantitative real-time PCR (qRT-PCR) and immunohistochemistry assays. In vitro cell proliferation was tested through MTT assay and flow cytometer was used to detected Cell apoptosis. The activation of ERK(extracellular signal regulated kinase) was explored by Western Blot. Results In the present research, we found that the expression of RNF6 was high in both primary tissues and cervical cancer cells. RNF6 could promote cervical cancer HeLa cells growth. Once knockdown of RNF6 in cervical cancer cells, cell proliferation could be suppressed and cell apoptosis was promoted. Moreover, its elevation had an adverse effect on the prognosis of cervical cancer. Further studies showed that ERK activation is one of the potential mechanisms. Conclusion These findings provided evidence that the up-regulated RNF6 could activate the MAPK/ERK pathway to regulate the cell growth in cervical cancer, which suggested that RNF6 could be a promising target for diagnosis and treatment for cervical cancer.

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The Potential Oncogenic and MLN4924-Resistant Effects of CSN5 on Cervical Cancer Cells

10.21203/rs.3.rs-550782/v1 ◽

2021 ◽

Author(s):

Huilin Zhang ◽

Ping He ◽

Qing Zhou ◽

Yan Lu ◽

Bingjian Lu

Keyword(s):

Cell Cycle ◽

Cervical Cancer ◽

Cell Proliferation ◽

Cancer Cells ◽

Cancer Cell ◽

Cervical Cancer Cell ◽

Cervical Cancers ◽

Cervical Cancer Cells

Abstract BackgroundsCSN5, a member of Cop9 signalosome, is essential for protein neddylation. It has been supposed to serve as an oncogene in some cancers. However, the role of CSN5 has not been investigated in cervical cancer yet.MethodsData from TCGA cohorts and GEO dataset was analyzed to examine the expression profile of CSN5 in cervical cancers. The role of CSN5 on cervical cancer cell proliferation was investigated in cervical cancer cell lines, Siha and Hela, through CSN5 knockdown via CRISPR-CAS9. Western blot was used to detect the effect of CSN5 knockdown and overexpression. CCK8, clone formation assay and cell cycle assay were also employed. Besides, the role CSN5 knockdown in vivo was evaluated by xenograft tumor model. Moreover, MLN4924 was applied in Siha and Hela with CSN5 overexpression.ResultsWe found that downregulation of CSN5 in Siha and Hela cells inhibited cell proliferation in vitro and in vivo, and the inhibitory effects were largely rescued by CSN5 overexpression. Moreover, deletion of CSN5 caused cell cycle arrest rather than inducing apoptosis. Importantly, CSN5 overexpression confers resistance to the anti-cancer effects of MLN4924 (pevonedistat) in cervical cancer cells.ConclusionsOur findings demonstrated that CSN5 functions as an oncogene in cervical cancers and may serve as a potential indicator for predicting the effects of MLN4924 treatment in the future.

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Hsp90 Inhibitor SNX-2112 Enhances TRAIL-Induced Apoptosis of Human Cervical Cancer Cells via the ROS-Mediated JNK-p53-Autophagy-DR5 Pathway

Oxidative Medicine and Cellular Longevity ◽

10.1155/2019/9675450 ◽

2019 ◽

Vol 2019 ◽

pp. 1-26

Author(s):

Liubing Hu ◽

Yan Wang ◽

Zui Chen ◽

Liangshun Fu ◽

Sheng Wang ◽

...

Keyword(s):

Cervical Cancer ◽

Cancer Cells ◽

Signaling Pathway ◽

Hela Cells ◽

Cell Apoptosis ◽

Hsp90 Inhibitor ◽

Cervical Cancer Cells ◽

Ros Scavenger ◽

Induced Apoptosis ◽

Human Cervical Cancer

Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) is a potent cancer cell apoptosis-inducing factor that can induce apoptosis in a variety of cancer cells. However, resistance to TRAIL in cancer cells is a huge obstacle in creating effective TRAIL-targeted clinical therapies. Thus, agents that can either enhance the effect of TRAIL or overcome its resistance are needed. In this study, we combined TRAIL with SNX-2112, an Hsp90 inhibitor we previously developed, to explore the effect and mechanism that SNX-2112 enhanced TRAIL-induced apoptosis in cervical cancer cells. Our results showed that SNX-2112 markedly enhanced TRAIL-induced cytotoxicity in HeLa cells, and this combination was found to be synergistic. Additionally, we found that SNX-2112 sensitized TRAIL-mediated apoptosis caspase-dependently in TRAIL-resistant HeLa cells. Mechanismly, SNX-2112 downregulated antiapoptosis proteins, including Bcl-2, Bcl-XL, and FLIP, promoted the accumulation of reactive oxygen species (ROS), and increased the expression levels of p-JNK and p53. ROS scavenger NAC rescued SNX-2112/TRAIL-induced apoptosis and suppressed SNX-2112-induced p-JNK and p53. Moreover, SNX-2112 induced the upregulation of death-receptor DR5 in HeLa cells. The silencing of DR5 by siRNA significantly decreased cell apoptosis by the combined effect of SNX-2112 and TRAIL. In addition, SNX-2112 inhibited the Akt/mTOR signaling pathway and induced autophagy in HeLa cells. The blockage of autophagy by bafilomycin A1 or Atg7 siRNA abolished SNX-2112-induced upregulation of DR5. Meanwhile, ROS scavenger NAC, JNK inhibitor SP600125, and p53 inhibitor PFTα were used to verify that autophagy-mediated upregulation of DR5 was regulated by the SNX-2112-stimulated activation of the ROS-JNK-p53 signaling pathway. Thus, the combination of SNX-2112 and TRAIL may provide a novel strategy for the treatment of human cervical cancer by overcoming cellular mechanisms of apoptosis resistance.

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Swertiamarin exerts anticancer effects on human cervical cancer cells via induction of apoptosis, inhibition of cell migration and targeting of MEK-ERK pathway

Tropical Journal of Pharmaceutical Research ◽

10.4314/tjpr.v20i1.12 ◽

2021 ◽

Vol 20 (1) ◽

pp. 75-81

Author(s):

Xinxiang Wang ◽

Tao Wang

Keyword(s):

Cervical Cancer ◽

Cancer Cells ◽

Western Blotting ◽

Hela Cells ◽

Migration And Invasion ◽

Inhibition Of Proliferation ◽

Anticancer Effects ◽

Cervical Cancer Cells ◽

Human Cervical Cancer

Purpose: To investigate the anticancer effects of swertiamarin against taxol- resistant human cervical cancer cells.Method: Cell viability was investigated using 3-(4,5-dimethylthiazol-2-yl)-2,5–diphenyl tetrazolium bromide (MTT) assay while colony survival was evaluated by clonogenic assay. Apoptotic cell death was assessed by AO/ETBR staining and western blotting techniques. The levels of reactive oxygen species (ROS) were measured using 2,7, dicholoro dihydrofluoresceindiacetate (H2DCFDA) staining.Cell migration and invasion were monitored with Transwell chamber assay. Western blotting assay was used to determine the expression levels of proteins of the MEK/ERK signaling pathway.Results: Swertiamarin induced dose- and time-dependent inhibition of proliferation of HeLa cervical cancer cells (p < 0.05). It also suppressed the colony formation potential of HeLa cells, and induced various structural modifications in HeLa cells. Swertiamarin exposure resulted in the formation of earlyapoptotic, late-apoptotic and necrotic cells, and significant modulation of apoptosis-allied proteins. It was observed that the migration and invasion of HeLa cells were potentially suppressed in dose-reliant fashion by swertiamarin. Western blotting results showed that the expressions of p-MEK and p-ERK were markedly reduced, while those of MEK and ERK were unaffected (p < 0.05).Conclusion: Swertiamarin exerts in vitro anticancer activity against cervical cancer cells (HeLa). Thus, it is promising for use in cervical cancer chemotherapy. However, there is need for confirmation of these findings through further in vivo and in vitro investigations. Keywords: Swertiamarin, Gentianaceae, Triterpene Sapogenin, Cervical cance

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Oleanolic acid inhibits the proliferation of Hela cells in cervical cancer by regulating the ACSL4 ferroptosis signaling pathway

10.21203/rs.3.rs-102345/v1 ◽

2020 ◽

Author(s):

Xiaofei Jiang ◽

Mingqing Shi ◽

Miao Sui ◽

Yizhen Yuan ◽

Shuang Zhang ◽

...

Keyword(s):

Cervical Cancer ◽

Hela Cell ◽

Cancer Cells ◽

Oleanolic Acid ◽

Hela Cells ◽

Proliferation Capacity ◽

Cervical Cancer Cells ◽

Related Proteins

Abstract Background: Cervical cancer continues to be the leading cause of cancer deaths among women worldwide. Oleanolic acid (OA) is a naturally occurring substance found in the leaves, fruits, and rhizomes of plants that has anti-cancer activity. Methods: We used tumor-bearing mice as the animal model and Hela cell as cell models. Western blot was used for detecting the expression of proteins in ferroptosis related proteins acyl-CoA synthase long-chain family member 4 (ACSL4), ferritin heavy chain (FTH1), transferrin receptor (TfR1) and glutathione peroxidase 4 (GPX4) in vivo and in vitro. MTT and EdU was for the detection of the viability of Hela cells. Results: In vivo experiments showed that OA significantly reduced the size and mass of cervical cancer tumors. In vitro experiments showed that OA significantly reduced the viability and proliferation capacity of Hela cells. In both in vivo and in vitro assays, OA increased the level of oxidative stress and Fe2+ content, and increased the expression of ferroptosis related proteins. We found high expression of ACSL4 in both xenograft models and cervical carcinoma cells. Meanwhile, knockdown of ACSL4 expression using shRNA in cervical cancer cells significantly increased cell viability and proliferation. In addition, decreased ROS levels and GPX4 were detected in ACSL4 knockdown cervical cancer cells, suggesting that ACSL4 inhibition may contribute to the reduction of ferroptosis within Hela cells and thus improve Hela cell survival. Conclusion: Promotion of ACSL4 dependent ferroptosis through OA may be an effective approach to treat cervical cancer.

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HOTAIR contributes to cell proliferation and metastasis of cervical cancer via targetting miR-23b/MAPK1 axis

Bioscience Reports ◽

10.1042/bsr20171563 ◽

2018 ◽

Vol 38 (1) ◽

Cited By ~ 14

Author(s):

Qin Li ◽

Yanhong Feng ◽

Xu Chao ◽

Shuai Shi ◽

Man Liang ◽

...

Keyword(s):

Cervical Cancer ◽

Cell Proliferation ◽

Cancer Cells ◽

Mitogen Activated Protein Kinase ◽

Cancer Tissue ◽

Tissue Samples ◽

Downstream Target ◽

Cervical Cancer Cells

The long non-coding RNA (lncRNA) HOX transcript antisense RNA (HOTAIR) has been found to be overexpressed in many human malignancies and involved in tumor progression and metastasis. Although the downstream target through which HOTAIR modulates tumor metastasis is not well-known, evidence suggests that miR-23b might be involved in this event. In the present study, the expressions of HOTAIR and miR-23b were detected by real-time PCR in 33 paired cervical cancer tissue samples and cervical cell lines. The effects of HOTAIR on the expressions of miR-23b and mitogen-activated protein kinase 1 (MAPK1) were studied by overexpression and RNAi approaches. We found that HOTAIR expression was significantly increased in cervical cancer cells and tissues. In contrast, the expression of miR-23b was obviously decreased. We further demonstrated that HOTAIR knockdown promoted apoptosis and inhibited cell proliferation and invasion in vitro and in vivo. Moreover, our data indicated that HOTAIR may competitively bind miR-23b and modulate the expression of MAPK1 indirectly in cervical cancer cells. Taken together, our study has identified a novel pathway through which HOTAIR exerts its oncogenic role, and provided a molecular basis for potential applications of HOTAIR in the prognosis and treatment of cervical cancer.

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Assessment of Cytotoxic Activity of Rosemary (Rosmarinus officinalisL.), Turmeric (Curcuma longaL.), and Ginger (Zingiber officinaleR.) Essential Oils in Cervical Cancer Cells (HeLa)

The Scientific World JOURNAL ◽

10.1155/2016/9273078 ◽

2016 ◽

Vol 2016 ◽

pp. 1-8 ◽

Cited By ~ 7

Author(s):

P. A. S. R. Santos ◽

G. B. Avanço ◽

S. B. Nerilo ◽

R. I. A. Marcelino ◽

V. Janeiro ◽

...

Keyword(s):

Cervical Cancer ◽

Cell Death ◽

Essential Oils ◽

Cytotoxic Activity ◽

Cancer Cells ◽

Hela Cells ◽

Membrane Integrity ◽

Annexin V ◽

Cervical Cancer Cells

The objective of this study was to evaluate the cytotoxic activity of rosemary (REO,Rosmarinus officinalisL.), turmeric (CEO,Curcuma longaL.), and ginger (GEO,Zingiber officinaleR.) essential oils in HeLa cells. Cytotoxicity tests were performedin vitro, using tetrazolium (MTT) and neutral red assays for evaluation of antiproliferative activity by different mechanisms, trypan blue assay to assess cell viability and evaluation of cell morphology for Giemsa to observe the cell damage, and Annexin V to evaluate cell death by apoptosis. CEO and GEO exhibited potent cytotoxic activity against HeLa cells. IC50obtained was 36.6 μg/mL for CEO and 129.9 μg/mL for GEO. The morphology of HeLa cells showed condensation of chromatin, loss of cell membrane integrity with protrusions (blebs), and cell content leakage for cells treated with CEO and GEO, from the lowest concentrations studied, 32.81 μg/mL of CEO and 32.12 μg/mL of GEO. The Annexin V assay revealed a profile of cell death by apoptosis for both CEO and GEO. The results indicate cytotoxic activityin vitrofor CEO and GEO, suggesting potential use as anticancer agents for cervical cancer cells.

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Extracellular Vesicles Long Non-Coding RNA AGAP2-AS1 Contributes to Cervical Cancer Cell Proliferation Through Regulating the miR-3064-5p/SIRT1 Axis

Frontiers in Oncology ◽

10.3389/fonc.2021.684477 ◽

2021 ◽

Vol 11 ◽

Author(s):

Min Li ◽

Jing Wang ◽

Hongli Ma ◽

Li Gao ◽

Kunxiang Zhao ◽

...

Keyword(s):

Cervical Cancer ◽

Cell Proliferation ◽

Cancer Cells ◽

Cancer Cell ◽

Cancer Progression ◽

Molecular Mechanisms ◽

Cervical Cancer Cell ◽

Cancer Cell Proliferation ◽

Cervical Cancer Cells

Cervical cancer is one of the most severe and prevalent female malignancies and a global health issue. The molecular mechanisms underlying cervical cancer development are poorly investigated. As a type of extracellular membrane vesicles, EVs from cancer cells are involved in cancer progression by delivering regulatory factors, such as proteins, microRNAs (miRNAs), and long non-coding RNAs (lncRNAs). In this study, we identified an innovative function of extracellular vesicle (EV) lncRNA AGAP2-AS1 in regulating cervical cancer cell proliferation. The EVs were isolated from the cervical cancer cells and were observed by transmission electron microscopy (TEM) and were confirmed by analyzing exosome markers. The depletion of AGAP2-AS1 by siRNA significantly reduced its expression in the exosomes from cervical cancer and in the cervical cancer treated with AGAP2-AS1-knockdown exosomes. The expression of AGAP2-AS1 was elevated in the clinical cervical cancer tissues compared with the adjacent normal tissues. The depletion of EV AGAP2-AS1 reduced cell viabilities and Edu-positive cervical cancer cells, while it enhanced cervical cancer cell apoptosis. Tumorigenicity analysis in nude mice showed that the silencing of EV AGAP2-AS1 attenuated cervical cancer cell growth in vivo. Regarding the mechanism, we identified that AGAP2-AS1 increased SIRT1 expression by sponging miR-3064-5p in cervical cancer cells. The overexpression of SIRT1 or the inhibition of miR-3064-5p reversed EV AGAP2-AS1 depletion-inhibited cancer cell proliferation in vitro. Consequently, we concluded that EV lncRNA AGAP2-AS1 contributed to cervical cancer cell proliferation through regulating the miR-3064-5p/SIRT1 axis. The clinical values of EV lncRNA AGAP2-AS1 and miR-3064-5p deserve to be explored in cervical cancer diagnosis and treatments.

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Anti-Cancer Effects of miR-181/PTEN/PI3K/Akt Signaling Pathway Suppressed Cell Proliferation and Migration of Human Cervical Cancer

Journal of Biomaterials and Tissue Engineering ◽

10.1166/jbt.2020.2311 ◽

2020 ◽

Vol 10 (5) ◽

pp. 615-623

Author(s):

Xiuling Wang ◽

Lan Luo ◽

Lina Xu ◽

Yumin Chen ◽

Xiaofei Liu ◽

...

Keyword(s):

Cervical Cancer ◽

Cell Proliferation ◽

Cancer Cells ◽

Cell Apoptosis ◽

Antitumor Effect ◽

Akt Signaling ◽

Akt Signaling Pathway ◽

Protein Levels ◽

Ldh Activity ◽

Cervical Cancer Cells

Cervical cancer ranks the second in the incidence of common women malignant tumors, which is the commonest malignant tumor in women's reproductive organs. In present research, we evaluated the possible antitumor effect of miR-181 on cervical cancer and explored the molecular mechanism of this phenomenon. Briefly, MTT, LDH activity kit and flow cytometry were used to detect cell growth, LDH activity and cell apoptosis rate of HeLa cells. The experiment correlation protein expression levels of PTEN, phosphatidylinositol-3-hydroxykinase (PI3K) and phosphorylation (p)-Akt (protein kinase B) were measured by Western blot analysis. In present research, this research observed that miR-181 weakened the cell proliferation activity of cervical cancer cells and induced their caspase3/9 to promotion of cell apoptosis. PTEN expression was induced at the genetic and protein levels, and the PI3K/Akt expressions were inhibited at the protein levels in cervical cancer cells. SiPTEN weakened the antitumor effect of miR-181 on cervical cancer by activation of PI3K/Akt signaling pathway.

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Effectiveness of Pleurotus ostreatus Extract Through Cytotoxic Test and Apoptosis Mechanism of Cervical Cancer Cells

Biosaintifika Journal of Biology & Biology Education ◽

10.15294/biosaintifika.v9i1.7546 ◽

2017 ◽

Vol 9 (1) ◽

pp. 148

Author(s):

Nuraeni Ekowati ◽

Aris Mumpuni ◽

Juni Safitri Muljowati

Keyword(s):

Cervical Cancer ◽

Cell Proliferation ◽

Cell Death ◽

Hela Cell ◽

Cancer Cells ◽

Bioactive Compounds ◽

Hela Cells ◽

Pleurotus Ostreatus ◽

Proliferation Kinetics ◽

Cervical Cancer Cells

Pleurotus ostreatus is a common mushroom cultivated in Indonesia, and potential properties of bioactive compounds for medicinal mushroom. This study was aimed at obtaining P.ostreatus extract bioactive compounds potential in inhibiting the proliferation of cervical cancer cells (HeLa) and evaluating the HeLa cell proliferation kinetics and HeLa cell death mechanisms. The research was beneficial in making this product can be easily applied in a more controlled industrial scale. Anticancer activity test through a cytotoxic test using the MTT [3- (4,5-dimetiltiazol-2-yl) -2.5-diphenyl tertrazolium bromide], the kinetics proliferation of HeLa cells and HeLa cell death mechanism was performed. Linear regression analysis was used to analyze the data. Ethyl acetate extract of P. ostreatus isolated from Madiun showed the best results with IC 50 = 107.59 µg / ml. HeLa cell proliferation kinetics analysis showed that the application of bioactive compounds 100 µg / ml resulted in an increase of in death of HeLa cells along with length of incubation time. An important finding was that HeLa cells death by apoptosis was greater than by necrosis. In conclusion, the extracts of P. ostreatus has the potential to inhibit the growth of HeLa cells.

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Synthesis and Cytotoxic Activity of Salicyloyl Hydrazone Derivatives

Applied Mechanics and Materials ◽

10.4028/www.scientific.net/amm.320.522 ◽

2013 ◽

Vol 320 ◽

pp. 522-525

Author(s):

Xiao Yang Qiu ◽

An Ran Shi ◽

Xiao Li Zhang

Keyword(s):

Cervical Cancer ◽

Melting Point ◽

Cytotoxic Activity ◽

Cancer Cells ◽

Hela Cells ◽

Elemental Analyses ◽

Cervical Cancer Cells ◽

Human Cervical Cancer Cells ◽

Human Cervical Cancer

Three salicyloyl hydrazone derivatives (compounds 1-3) were prepared by reacting salicyloyl hydrazine with substituted formaldehydes. Their structures were characterized by melting point, 1H-NMR, ESI-MS and elemental analyses. The cytotoxic activity of compounds 1-3 was evaluated in vitro against Hela cells (human cervical cancer cells). The results revealed that all the compounds showed cytotoxic activity, with IC50 values lower than 15 μM.

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