Downregulation of HBx Restrains Proliferation, Migration, and Invasion of HepG2 Cells

Liver cancer is a major contributor to cancer-related death with poor survival for sufferers. Meanwhile, Hepatic B virus X protein (HBx) and XB130 are likely to participate in the pathogenesis of liver cancer. However, the detailed mechanism of HBx/XB130 in liver cancer remains to be further investigated. Our study explored the effects of HBx/XB130 on liver cancer progression. HBx and XB130 expression was detected by reverse transcription quantitative polymerase chain reaction (RT-qPCR) and Western blot. Overexpression of HBx and XB130 was found in liver cancer tissues and cells. Mechanistic study revealed that HBx could bind to and positively regulate XB130 in HepG2 cells. Subsequently, HBx expression was knocked down, while XB130 was overexpressed in HepG2 cells in order to observe the specific role of HBx/XB130 in liver cancer in vitro. Results of CCK-8, Transwell, wound healing, and colony formation assays suggested that HBx could mediate biological function of HepG2 cells by activating the XB130-mediated PI3K/AKT pathway. In summary, our data illustrate that inhibition of HBx effectively suppressed proliferation and metastasis and induced apoptosis of liver cancer cells, which might be partially reversed by XB130. HBx and XB130 may be potential targets for liver cancer pathogenesis.

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Downregulation of HBx Restrains Proliferation, Migration and Invasion of HepG2 Cells

10.21203/rs.3.rs-36398/v1 ◽

2020 ◽

Author(s):

Chaoqun Huang ◽

Wei Liu ◽

Xiaochuan Zhao ◽

Libin Zhao ◽

Fuxiang Wang

Keyword(s):

Liver Cancer ◽

Colony Formation ◽

Hepg2 Cells ◽

Cellular Proliferation ◽

Quantitative Polymerase Chain Reaction ◽

Migration And Invasion ◽

Loss Of Function ◽

Normal Tissues ◽

Cancer Tissues ◽

Rip Assay

Abstract Background: Liver cancer is a frequent malignancy with high fatality. Hepatic B virus X protein (HBx) could promote theprogression of liver cancer. Meanwhile, aberrantly expressed XB130 was identified in liver cancer. However, relevant molecular mechanism is poorly studied. Our present study mainly investigated the mechanism of liver cancer.Methods: After microarray-based analyses in liver cancer tissues and the matched adjacent normal tissues, upregulated mRNA was screened out. Contents of HBx and XB130 in liver cancer tissues as well as cells HepG2 were examined by reverse transcription quantitative polymerase chain reaction (RT-qPCR) and Western blot analysis. Correlation between HBx and XB130 was analyzed in HepG2 cells, followed by verification by RIP assay. After gain- and loss-of-function experiments, cellular proliferation, invasion, migration and colony formation ability were assessed using CCK-8, Transwell, wound healing experiment and colony formation assay.Results: Both HBx and XB130 expression was elevated in liver cancer, which was correlated with poor survival rate of liver cancer patients. Moreover, HBx and XB130 were positively correlated in HepG2 cells. RIP assay verified that HBx could bind to XB130. Loss of HBx hindered proliferation, and migration/invasion of HepG2 cells but promoted apoptosis, which was partially reversed by overexpressed XB130.Conclusion: The conclusion reached from the study offered an understanding of the role of HBx/XB130 played in liver cancer. Our study first reported the regulatory relation between HBx and XB130, which may be valuable to discover therapeutic targets for liver cancer treatment.

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miR-183-5p promotes proliferation and migration in hepatocellular carcinoma by targeting IRS1 and its association with patient survival

The International Journal of Biological Markers ◽

10.1177/1724600820951572 ◽

2020 ◽

Vol 35 (3) ◽

pp. 83-89

Author(s):

Rong Yan ◽

Kang Li ◽

Dawei Yuan ◽

Haonan Wang ◽

Wei Chen ◽

...

Keyword(s):

Hepatocellular Carcinoma ◽

Liver Cancer ◽

Cancer Progression ◽

Normal Tissues ◽

Report Analysis ◽

In Vitro Effects ◽

And Migration ◽

Cancer Tissues ◽

Proliferation And Migration

Background: MiR-183-5p plays an important role in the pathophysiology of many tumors, while the role of MiR-183-5p in liver cancer is unclear. Methods: In this study, quantitative reverse transcription-polymerase chain reaction and Western blotting were used to detect the expression of miR-183-5p in liver cancer cell lines, liver cancer tissues, and normal tissues adjacent to the cancer, and to explore the mechanism of miR-183-5p regulating liver cancer progression. The in vitro effects of miR-183-5p were evaluated by CCK-8, colony formation test, and wound healing test. Various databases were used to predict the target mRNA of miR-183-5p and verified by luciferase report analysis. In addition, the effects of miR-183-5p and its target gene on the survival of patients with liver cancer were also analyzed. Results: miR-183-5p was highly expressed in hepatocellular carcinoma cells and tissues, and was related to some clinicopathological features. MiR-183-5p can promote the proliferation and migration of liver cancer cells. Using the bioinformatics database, we proved that miR-183-5p is related to the survival of liver cancer patients. Insulin receptor substrate 1 (IRS1) is a target of miR-183-5p, and luciferase analysis confirmed that miR-183-5p combines with the 3′-untranslated region (3′-UTR) of IRS1. Conclusion: The miR-183-5p/IRS1 axis may be a new target for liver cancer research.

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USP44 hypermethylation promotes cell proliferation and metastasis in breast cancer

Future Oncology ◽

10.2217/fon-2020-0415 ◽

2020 ◽

Author(s):

Xin Chen ◽

Xiaotang Wu ◽

Wen Lei

Keyword(s):

Breast Cancer ◽

Cell Proliferation ◽

Cancer Progression ◽

Migration And Invasion ◽

Expression Levels ◽

Cell Behaviors ◽

Specific Pcr ◽

Proliferation And Metastasis ◽

Induced Apoptosis

Aim: The methylation and expression levels of USP44 in breast cancer were investigated and their effects on tumor cells were researched. Materials & Methods: Bioinformatics was employed to identify the target gene from TCGA database. Sodium bisulfite and decitabine were used for DNA modification and demethylation, and methylation-specific PCR and reverse transcriptase PCR were performed to assess USP44 methylation and expression levels. Tumor cell behaviors were assayed via several in vitro experiments. Results: USP44 was hypermethylated, which caused its poor expression in breast cancer, whereas its overexpression significantly suppressed cancer cell proliferation, migration and invasion and induced apoptosis. Conclusion: USP44 negatively functions in cancer progression upon overexpression, indicating its potential as a therapeutic target for clinical treatment of breast cancer.

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circSLC6A6 Sponges miR-497-5p to Promote Endometrial Cancer Progression via the PI4KB/Hedgehog Axis

Journal of Immunology Research ◽

10.1155/2021/5512391 ◽

2021 ◽

Vol 2021 ◽

pp. 1-13

Author(s):

Juan Hu ◽

Xing Peng ◽

Weina Du ◽

Yichuan Huang ◽

Chun Zhang ◽

...

Keyword(s):

Endometrial Cancer ◽

Cancer Progression ◽

Hedgehog Pathway ◽

Luciferase Reporter ◽

Mechanistic Study ◽

Migration And Invasion ◽

Circular Rnas ◽

Ec Cells

Background. As a new kind of noncoding RNAs, circular RNAs (circRNAs) have been substantiated to be involved in multiple biological processes. Accumulating studies indicate that circular RNAs (circRNAs) regulate the development of cancers by acting as miRNA sponges. However, the role of circRNAs in endometrial cancer (EC) is rarely reported. This study was aimed at investigating the functional roles of circSLC6A6 in EC. Methods. The qRT-PCR assay was performed to detect the circSLC6A6 expression in EC tissues and cell lines. The luciferase reporter assay was performed to explore the connection between circSLC6A6 and miR-497-5p as well as the connection between miR-497-5p and PI4KB. The colony formation assay, EdU assay, wound healing assay, and transwell assay were performed to examine the proliferation, migration, and invasion of EC cells. The in vivo assay was performed to reveal the function of circSLC6A6 in tumorigenesis. Results. We found that circSLC6A6 was highly expressed in both EC tissues and cells. And circSLC6A6 promoted the proliferation, migration, and invasion of EC cells in vitro. In vivo, circSLC6A6 promoted tumor growth. Besides, a mechanistic study demonstrated that circSLC6A6 could regulate tumor-associated signaling PI4KB/hedgehog pathway by sponging miR-497-5p. Conclusion. This study illustrates that circSLC6A6 plays a role in promoting EC progression via the miR-497-5p-mediated PI4KB/hedgehog pathway. Our study may provide a potential novel biomarker for EC diagnosis or treatment.

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SPINT1-AS1 Drives Cervical Cancer Progression via Repressing miR-214 Biogenesis

Frontiers in Cell and Developmental Biology ◽

10.3389/fcell.2021.691140 ◽

2021 ◽

Vol 9 ◽

Author(s):

Hongjuan Song ◽

Yuan Liu ◽

Hui Liang ◽

Xin Jin ◽

Liping Liu

Keyword(s):

Cervical Cancer ◽

Cancer Progression ◽

Cellular Proliferation ◽

Migration And Invasion ◽

Downstream Target ◽

Non Coding Rna ◽

Non Coding Rnas ◽

Cancer Tissues

Accumulating evidences have revealed the dysregulated expressions and critical roles of non-coding RNAs in various malignancies, including cervical cancer. Nevertheless, our knowledge about the vast majority of non-coding RNAs is still lacking. Here we identified long non-coding RNA (lncRNA) SPINT1-AS1 as a novel cervical cancer-associated lncRNA. SPINT1-AS1 was increased in cervical cancer and correlated with advanced stage and poor prognosis. SPINT1-AS1 was a direct downstream target of miR-214, a well-known tumor suppressive microRNA (miRNA) in cervical cancer. Intriguingly, SPINT1-AS1 was also found to repress miR-214 biogenesis via binding DNM3OS, the primary transcript of miR-214. The interaction between SPINT1-AS1 and DNM3OS repressed the binding of DROSHA and DGCR8 to DNM3OS, blocked DNM3OS cleavage, and therefore repressed mature miR-214 biogenesis. The expression of SPINT1-AS1 was significantly negatively correlated with miR-214 in cervical cancer tissues, supporting the reciprocal repression between SPINT1-AS1 and miR-214 in vivo. Through downregulating mature miR-214 level, SPINT1-AS1 upregulated the expression of β-catenin, a target of miR-214. Thus, SPINT1-AS1 further activated Wnt/β-catenin signaling in cervical cancer. Functionally, SPINT1-AS1 drove cervical cancer cellular proliferation, migration, and invasion in vitro, and also tumorigenesis in vivo. Deletion of the region mediating the interaction between SPINT1-AS1 and DNM3OS, overexpression of miR-214, and inhibition of Wnt/β-catenin signaling all reversed the roles of SPINT1-AS1 in cervical cancer. Collectively, these findings identified SPINT1-AS1 as a novel cervical cancer-associated oncogenic lncRNA which represses miR-214 biogenesis and activates Wnt/β-catenin signaling, highlighting its potential as prognostic biomarker and therapeutic target for cervical cancer.

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miR-1-3p Contributes to Cell Proliferation and Invasion by Targeting Glutaminase in Bladder Cancer Cells

Cellular Physiology and Biochemistry ◽

10.1159/000495273 ◽

2018 ◽

Vol 51 (2) ◽

pp. 513-527 ◽

Cited By ~ 9

Author(s):

Junfeng Zhang ◽

Longsheng Wang ◽

Shiyu Mao ◽

Mengnan Liu ◽

Wentao Zhang ◽

...

Keyword(s):

Cell Proliferation ◽

Bladder Cancer ◽

Cancer Cells ◽

Cancer Progression ◽

Methylation Status ◽

Luciferase Reporter ◽

Migration And Invasion ◽

Bladder Cancer Cells ◽

Cancer Tissues

Background/Aims: Increasing evidence showed that miR-1-3p plays a major role in malignant tumor progression. However, the specific biological function of miR-1-3p in bladder cancer is yet unknown. Methods: The expression levels of miR-1-3p in bladder cancer tissues and cell lines were examined by qRT-PCR. Bisulfite sequencing PCR was used for DNA methylation analysis. The target of miR-1-3p was validated by a dual luciferase reporter assay, and the effects of miR-1-3p on phenotypic changes in bladder cancer cells were investigated in vitro and in vivo. Results: The expression of miR-1-3p in bladder cancer cells was downregulated as compared to normal SV-HUC-1 cells. Also, the expression of miR-1-3p was significantly lower in bladder cancer tissues than the corresponding non-cancerous tissues. The methylation status of CpG islands was involved in the regulation of miR-1-3p expression. miR-1-3p inhibited the bladder cancer cell proliferation, migration, and invasion by directly targeting the 3’-UTR of glutaminase. It also exerted an anti-tumor effect by negatively regulating the glutaminase in a xenograft mouse model. Furthermore, GLS depletion resulted in the prolonged expression of γH2AX. Conclusion: Taken together, these results demonstrated that miR-1-3p acts as a tumor suppressor via regulation of glutaminase expression in bladder cancer progression, and miR-1-3p might represent a novel therapeutic target for the treatment of bladder cancer.

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MicroRNA-937 is overexpressed and predicts poor prognosis in patients with colon cancer

Diagnostic Pathology ◽

10.1186/s13000-019-0920-3 ◽

2019 ◽

Vol 14 (1) ◽

Cited By ~ 1

Author(s):

Huiya Liu ◽

Lin Ma ◽

Ling Wang ◽

Yizuo Yang

Keyword(s):

Colon Cancer ◽

Cell Proliferation ◽

Cell Lines ◽

Cox Regression ◽

Quantitative Polymerase Chain Reaction ◽

Cell Counting ◽

Migration And Invasion ◽

Prognostic Impact ◽

Cancer Tissues

Abstract Background Colon cancer is a heterogeneous tumor and a leading cause of cancer-related mortality. MicroRNA (miRNA) has been proposed as the biomarker in cancers. The aim of this study was to investigate the clinical significance and potential functional role of miR-937 in colon cancer. Methods In the present study, reverse transcription-quantitative polymerase chain reaction (qRT-PCR) was conducted to examine the expression levels of miR-937 in colon cancer tissues and cell lines. Kaplan-Meier curve and Cox regression analyses were used to determine the prognostic impact of miR-937 on survival. Cell Counting Kit-8 and Transwell assays were performed to examine cell proliferation, migration, and invasion, respectively. Results miR-937 was significantly upregulated in colon cancer tissues and cell lines. Clinical analysis results showed that miR-937 expression was associated with lymph node metastasis and TNM stage. Patients with high miR-937 expression predicted a shorter overall survival rate. Functionally, overexpression of miR-937 promoted cell proliferation, migration, and invasion, while inhibition of miR-937 inhibited these cellular behaviors in vitro. Conclusions These results suggested that miR-937 may act as a prognostic biomarker and a potential target for therapeutic strategy, as well as promote proliferation, migration, and invasion of colon cancer.

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Total Flavonoids of Litchi Seed Attenuate Prostate Cancer Progression Via Inhibiting AKT/mTOR and NF-kB Signaling Pathways

Frontiers in Pharmacology ◽

10.3389/fphar.2021.758219 ◽

2021 ◽

Vol 12 ◽

Author(s):

Ming Chang ◽

Dan Zhu ◽

Yanjiang Chen ◽

Weiquan Zhang ◽

Xi Liu ◽

...

Keyword(s):

Prostate Cancer ◽

Cell Proliferation ◽

Cell Growth ◽

Signaling Pathways ◽

Cancer Progression ◽

Molecular Mechanisms ◽

Migration And Invasion ◽

Induced Apoptosis

Litchi seeds have been traditionally used in Chinese herbal formula for urologic neoplasms including prostate cancer (PCa). However, the effective components of Litchi seeds and the mechanisms of their actions on PCa cell growth and metastasis remain unclear. In this study, we investigated the effects and molecular mechanisms of the Total Flavonoid of Litchi Seed (TFLS) in PCa PC3 and DU145 cell lines. We found that TFLS significantly inhibited the PCa cell proliferation, induced apoptosis, and prevented cell migration and invasion. Furthermore, we observed that TFLS upregulated the expression of epithelial biomarker E-cadherin and downregulated mesenchymal biomarker Vimentin. TFLS also increased the expression of cleaved-PRAP and Bax, and decreased the expression of Bcl-2 in both PC3 and DU145 cells. Besides, TFLS inhibited AKT signaling pathway by reducing the phosphorylation of AKT and activities of downstream signal transducers including mTOR, IκBα and NF-kB. Finally, TFLS treated mice exhibited a significant decrease in tumor size without toxicity in major organs in vivo. These results indicated that TFLS could suppress PCa cell growth in vivo and inhibit PCa cell proliferation and metastasis in vitro through induction of apoptosis and phenotypic reversal of EMT, which may be achieved by inhibiting the AKT/mTOR and NF-κB signaling pathways. Taken together, our data provide new insights into the role of TFLS as a novel potent anti-cancer agent for the treatment of PCa.

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Abnormal Expression of Centromere Protein U Is Associated with Hepatocellular Cancer Progression

BioMed Research International ◽

10.1155/2021/4051192 ◽

2021 ◽

Vol 2021 ◽

pp. 1-14

Author(s):

Yuanlin Yu ◽

Xiaopeng Chen ◽

Weidong Zhang ◽

Jun Liu

Keyword(s):

Gene Expression ◽

Cancer Progression ◽

Quantitative Polymerase Chain Reaction ◽

Gene Set Enrichment Analysis ◽

The Cancer Genome Atlas ◽

Cell Counting ◽

Migration And Invasion ◽

Abnormal Expression ◽

Centromere Protein

Background. Hepatocellular carcinoma (HCC) is one of the most common malignancies globally, but its molecular mechanism is unclear. Abnormal expression of centromere protein U (CENPU) is closely related to diverse human cancers. The purpose of this article was to evaluate the function and potential mechanisms of CENPU in HCC development. Methods. We performed bioinformatics analysis of The Cancer Genome Atlas (TCGA), Gene Expression Omnibus (GEO), Gene Expression Profiling Interactive Analysis (GEPIA), and Kaplan-Meier plotter databases to investigate the clinical significance and prognostic value of CENPU in HCC. Western blotting and immunohistochemical staining were used to measure protein expression, while reverse transcription-quantitative polymerase chain reaction (qRT-PCR) was used to determine mRNA expression. Cell Counting Kit8 (CCK-8) and colony formation assays were conducted to examine cell proliferation. Transwell and wound healing assays were used to assess cell migration and invasion. Gene set enrichment analysis (GSEA) was used to explore the potential signaling pathways of CENPU involved in HCC. Results. High expression of CENPU in HCC was predicted by public database analysis and indicated a poor prognosis. CENPU expression was significantly higher in HCC tissues and cells than in normal tissues and cell. In vitro, CENPU promoted the proliferation, migration, and invasion of HCC cells. GSEA results indicated that CENPU was linked to the Notch signaling pathway, and our research supported this prediction. Conclusion. CENPU promotes the malignant biological process of HCC and may be a promising target for HCC treatment.

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LncRNA XIST promotes liver cancer progression by acting as a molecular sponge of miR-200b-3p to regulate ZEB1/2 expression

Journal of International Medical Research ◽

10.1177/03000605211016211 ◽

2021 ◽

Vol 49 (5) ◽

pp. 030006052110162

Author(s):

Lili Liu ◽

Hua Jiang ◽

Hongming Pan ◽

Xiuming Zhu

Keyword(s):

Cell Proliferation ◽

Liver Cancer ◽

Cancer Progression ◽

Cancer Metastasis ◽

Cell Counting ◽

Luciferase Reporter ◽

Migration And Invasion ◽

Xist Expression ◽

Lncrna Xist

Objective To evaluate the predictive value of long non-coding RNA (lncRNA) X-inactive specific transcript (XIST) for survival, and determine the involvement of miRNA(miR)-200b-3p and zinc finger E-box-binding homeobox (ZEB) 1/2 in the pro-tumor effect of lncRNA XIST in liver cancer. Methods We evaluated lncRNA XIST expression in liver cancer tissues and cell lines by quantitative reverse transcription polymerase chain reaction (RT-qPCR) and analyzed the correlation between its expression and overall survival of liver cancer patients by Kaplan–Meier analysis. Its effects on cell proliferation, migration, and invasion were analyzed by Cell-Counting Kit-8 and Transwell assays. The association between lncRNA XIST and miR-200b-3p, and the effects of lncRNA XIST on ZEB1/2 expression were explored using luciferase reporter assays, real-time PCR, and western blotting. Results The lncRNA XIST was significantly upregulated in liver cancer, and increased lncRNA XIST expression was associated with a poor prognosis. The lncRNA XIST promoted liver cancer cell proliferation, migration, and invasion in vitro, and acted as a molecular sponge for miR-200b-3p, and also regulated the expression of ZEB1/2 via miR-200b-3p. Conclusion The lncRNA XIST is an oncogenic lncRNA that promotes liver cancer metastasis, and its pro-metastatic phenotype can be partially attributed to the lncRNA XIST/miR-200b-3p/ZEB1/2 signaling axis.

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