AIM To investigate the role of long noncoding RNA(lnc RNA) RP4 in colorectal cancer.METHODS Lentivirus-mediated lnc RNA RP4 overexpression and knockdown were performed in the colorectal cancer cell line SW480. Cell pr...AIM To investigate the role of long noncoding RNA(lnc RNA) RP4 in colorectal cancer.METHODS Lentivirus-mediated lnc RNA RP4 overexpression and knockdown were performed in the colorectal cancer cell line SW480. Cell proliferation, tumor growth, and early apoptosis were evaluated by a cell counting kit-8 assay, an in vivo xenograft tumor model, and annexin V/propidium iodide staining, respectively. Analysis of the lnc RNA RP4 mechanism involved assessment of the association of its expression with mi R-7-5 p and the SH3 GLB1 gene. Western blot analysis was also performed to assess the effect of lnc RNA RP4 on the autophagy-mediated cell death pathway and phosphatidylinositol-3-kinase(PI3 K)/Akt signaling.RESULTS Cell proliferation, tumor growth, and early apoptosis in SW480 cells were negatively regulated by lnc RNA RP4. Functional experiments indicated that lnc RNA RP4 directly upregulated SH3 GLB1 expression by acting as a competing endogenous RNA(ce RNA) for mi R-7-5 p. This interaction led to activation of the autophagy-mediated cell death pathway and de-repression of PI3 K and Akt phosphorylation in colorectal cancer cells in vivo.CONCLUSION Our results demonstrated that lnc RNA RP4 is a ce RNA that plays an important role in the pathogenesis of colorectal cancer, and could be a potential therapeutic target for colorectal cancer treatment.展开更多
Background:Drug resistance is the main factor contributing to cancer recurrence and poor prognosis.Exploration of drug resistance-related mechanisms and effective therapeutic targets are the aim of molecular targeted ...Background:Drug resistance is the main factor contributing to cancer recurrence and poor prognosis.Exploration of drug resistance-related mechanisms and effective therapeutic targets are the aim of molecular targeted therapy.In our study,the role of long non-coding RNA(lncRNA)AFAP1-AS1 in gemcitabine resistance and related mechanisms were explored in cervical cancer cells.Methods:Gemcitabine-resistant cervical cancer cell lines HT-3-Gem and SW756-Gem were constructed using the gemcitabine concentration gradient method.The overall survival rates and recurrence-free survival rates were evaluated by Kaplan-Meier analysis.The interaction was verified through a Dual-luciferase reporter gene assay and a Biotinylated RNA pull-down assay.Cell proliferation ability was assessed through methyl-thiazolyl-tetrazolium(MTT),soft agar,and colony formation experiments.Cell cycle and apoptosis were detected byflow cytometry.Results:Up-regulation of AFAP1-AS1 in cervical cancer predicted a poor prognosis.Besides,patients in the gemcitabine-resistance group had higher levels of AFAP1-AS1 than the gemcitabine-sensitive group.AFAP1-AS1 promoted tumor growth and induced gemcitabine tolerance of cervical cancer cells.In addition,AFAP1-AS1 mediated epidermal growth factor receptor(EGFR)expression by serving as a molecular sponge for microRNA-7a-5p(miR-7-5p).This present study also proved that the knockdown of EGFR or overexpression of miR-7a-5p abolished the accelerative role of AFAP1-AS1 overexpression in cancer progression and gemcitabine tolerance.Conclusions:In general,the AFAP1-AS1/miR-7-5p/EGFR axis was tightly related to the progression and gemcitabine tolerance of cervical cancer,providing potential targets for the management of cervical cancer.展开更多
Acidosis,regardless of hypoxia involvement,is recognized as a chronic and harsh tumor microenvironment(TME)that educates malignant cells to thrive and metastasize.Although overwhelming evidence supports an acidic envi...Acidosis,regardless of hypoxia involvement,is recognized as a chronic and harsh tumor microenvironment(TME)that educates malignant cells to thrive and metastasize.Although overwhelming evidence supports an acidic environment as a driver or ubiquitous hallmark of cancer progression,the unrevealed core mechanisms underlying the direct effect of acidification on tumorigenesis have hindered the discovery of novel therapeutic targets and clinical therapy.Here,chemical-induced and transgenic mouse models for colon,liver and lung cancer were established,respectively.miR-7 and TGF-β2 expressions were examined in clinical tissues(n=184).RNA-seq,miRNA-seq,proteomics,biosynthesis analyses and functional studies were performed to validate the mechanisms involved in the acidic TME-induced lung cancer metastasis.Our data show that lung cancer is sensitive to the increased acidification of TME,and acidic TME-induced lung cancer metastasis via inhibition of miR-7-5 p.TGF-β2 is a direct target of miR-7-5 p.The reduced expression of miR-7-5 p subsequently increases the expression of TGF-β2 which enhances the metastatic potential of the lung cancer.Indeed,overexpression of miR-7-5 p reduces the acidic p H-enhanced lung cancer metastasis.Furthermore,the human lung tumor samples also show a reduced miR-7-5 p expression but an elevated level of activated TGF-β2;the expressions of both miR-7-5 p and TGF-β2 are correlated with patients’survival.We are the first to identify the role of the miR-7/TGF-β2 axis in acidic p H-enhanced lung cancer metastasis.Our study not only delineates how acidification directly affects tumorigenesis,but also suggests miR-7 is a novel reliable biomarker for acidic TME and a novel therapeutic target for non-small cell lung cancer(NSCLC)treatment.Our study opens an avenue to explore the p H-sensitive subcellular components as novel therapeutic targets for cancer treatment.展开更多
Background and Aims:Intrahepatic cholangiocarcinoma(ICC)is a malignant tumor derived from intrahepatic bile duct epithelial cells.Accumulating studies report that microRNAs are widely involved in tumor migration and m...Background and Aims:Intrahepatic cholangiocarcinoma(ICC)is a malignant tumor derived from intrahepatic bile duct epithelial cells.Accumulating studies report that microRNAs are widely involved in tumor migration and metastasis by regulation of target genes.miR-7-5p has been confirmed to inhibit tumor metastasis and to be related to prognosis for several malignant tumors.Our study investigated the underlying functions of miR-7-5p in ICC.Methods:The expression of miR-7-5p in ICC tissues but also in ICC cell lines was analyzed by real-time PCR.By analyzing the relationship between the clinicopathological parameters of 60 ICC patients and the expression level of miR-7-5p,the effect of miR-7-5p on the prognosis was clarified.After transfected with miR-7-5p mimics or miR-7-5p inhibitor,cell counting kit-8 assay was applied to evaluate the cells proliferation,flow cytometry was applied to analyze the cells apoptosis,wound healing assay and transwell chamber assay were applied to analyze the cell invasion and migration.A luciferase reporter assay was identified the relationship of miR-7-5p and myeloid differentiation factor 88(MyD88).Western blotting was used to analyze the proteins expression.And immunochemistry was performed to determine the expression of MYD88 in ICC tissues.Results:Our data showed the expression of miR-7-5p was down-regulated not only in ICC tissues but also in ICC cell lines compared with normal controls.Low expression of miR-7-5p was notably associated with poor prognosis in ICC patients.miR-7-5p negatively regulated cell proliferation,migration,invasion and apoptosis in ICC cells.We further verified that MyD88 was a novel target of miR-7-5p and was significantly overexpressed in ICC tissues.Overexpression of MyD88 counteracted the effects of miR-7-5p in ICC cells.Conclusions:The present findings suggest that miR-7-5p plays a pivotal role in ICC invasion by regulating MyD88.Ampliative insight into the key factors of ICC invasion may result in the development of new treatment options for ICC.展开更多
基金Supported by Scientific Research Foundation of Anhui Education Department,No.KJ2017A219 to Liu MLScientific Research Foundation of Academic Leader of Anhui Province,No.2016H105 to Liu ML+2 种基金Education Talent Foundation of Universities of Anhui Education Department,No.gxbj ZD2016070 to Liu MLNational Natural Science Foundation of China,No.81500373 to Wang WBNatural Science Foundation of Anhui Province,No.1608085MH193 to Wang WB
文摘AIM To investigate the role of long noncoding RNA(lnc RNA) RP4 in colorectal cancer.METHODS Lentivirus-mediated lnc RNA RP4 overexpression and knockdown were performed in the colorectal cancer cell line SW480. Cell proliferation, tumor growth, and early apoptosis were evaluated by a cell counting kit-8 assay, an in vivo xenograft tumor model, and annexin V/propidium iodide staining, respectively. Analysis of the lnc RNA RP4 mechanism involved assessment of the association of its expression with mi R-7-5 p and the SH3 GLB1 gene. Western blot analysis was also performed to assess the effect of lnc RNA RP4 on the autophagy-mediated cell death pathway and phosphatidylinositol-3-kinase(PI3 K)/Akt signaling.RESULTS Cell proliferation, tumor growth, and early apoptosis in SW480 cells were negatively regulated by lnc RNA RP4. Functional experiments indicated that lnc RNA RP4 directly upregulated SH3 GLB1 expression by acting as a competing endogenous RNA(ce RNA) for mi R-7-5 p. This interaction led to activation of the autophagy-mediated cell death pathway and de-repression of PI3 K and Akt phosphorylation in colorectal cancer cells in vivo.CONCLUSION Our results demonstrated that lnc RNA RP4 is a ce RNA that plays an important role in the pathogenesis of colorectal cancer, and could be a potential therapeutic target for colorectal cancer treatment.
文摘Background:Drug resistance is the main factor contributing to cancer recurrence and poor prognosis.Exploration of drug resistance-related mechanisms and effective therapeutic targets are the aim of molecular targeted therapy.In our study,the role of long non-coding RNA(lncRNA)AFAP1-AS1 in gemcitabine resistance and related mechanisms were explored in cervical cancer cells.Methods:Gemcitabine-resistant cervical cancer cell lines HT-3-Gem and SW756-Gem were constructed using the gemcitabine concentration gradient method.The overall survival rates and recurrence-free survival rates were evaluated by Kaplan-Meier analysis.The interaction was verified through a Dual-luciferase reporter gene assay and a Biotinylated RNA pull-down assay.Cell proliferation ability was assessed through methyl-thiazolyl-tetrazolium(MTT),soft agar,and colony formation experiments.Cell cycle and apoptosis were detected byflow cytometry.Results:Up-regulation of AFAP1-AS1 in cervical cancer predicted a poor prognosis.Besides,patients in the gemcitabine-resistance group had higher levels of AFAP1-AS1 than the gemcitabine-sensitive group.AFAP1-AS1 promoted tumor growth and induced gemcitabine tolerance of cervical cancer cells.In addition,AFAP1-AS1 mediated epidermal growth factor receptor(EGFR)expression by serving as a molecular sponge for microRNA-7a-5p(miR-7-5p).This present study also proved that the knockdown of EGFR or overexpression of miR-7a-5p abolished the accelerative role of AFAP1-AS1 overexpression in cancer progression and gemcitabine tolerance.Conclusions:In general,the AFAP1-AS1/miR-7-5p/EGFR axis was tightly related to the progression and gemcitabine tolerance of cervical cancer,providing potential targets for the management of cervical cancer.
基金supported by the projects of National Natural Science Foundation of China(81874367 and 82074019)Guangdong Key Laboratory for Translational Cancer research of Chinese Medicine(2018B030322011,China)+3 种基金Natural Science Foundation for Distinguished Young Scholars of Guangdong Province,China(2017A030306033)Guangdong Province Universities and Colleges Pearl River Scholar Funded Scheme(2016,China)Project of Educational Commission of Guangdong Province of China(2016KTSCX012)Pearl River Nova Program of Guangzhou,China(201710010108)。
文摘Acidosis,regardless of hypoxia involvement,is recognized as a chronic and harsh tumor microenvironment(TME)that educates malignant cells to thrive and metastasize.Although overwhelming evidence supports an acidic environment as a driver or ubiquitous hallmark of cancer progression,the unrevealed core mechanisms underlying the direct effect of acidification on tumorigenesis have hindered the discovery of novel therapeutic targets and clinical therapy.Here,chemical-induced and transgenic mouse models for colon,liver and lung cancer were established,respectively.miR-7 and TGF-β2 expressions were examined in clinical tissues(n=184).RNA-seq,miRNA-seq,proteomics,biosynthesis analyses and functional studies were performed to validate the mechanisms involved in the acidic TME-induced lung cancer metastasis.Our data show that lung cancer is sensitive to the increased acidification of TME,and acidic TME-induced lung cancer metastasis via inhibition of miR-7-5 p.TGF-β2 is a direct target of miR-7-5 p.The reduced expression of miR-7-5 p subsequently increases the expression of TGF-β2 which enhances the metastatic potential of the lung cancer.Indeed,overexpression of miR-7-5 p reduces the acidic p H-enhanced lung cancer metastasis.Furthermore,the human lung tumor samples also show a reduced miR-7-5 p expression but an elevated level of activated TGF-β2;the expressions of both miR-7-5 p and TGF-β2 are correlated with patients’survival.We are the first to identify the role of the miR-7/TGF-β2 axis in acidic p H-enhanced lung cancer metastasis.Our study not only delineates how acidification directly affects tumorigenesis,but also suggests miR-7 is a novel reliable biomarker for acidic TME and a novel therapeutic target for non-small cell lung cancer(NSCLC)treatment.Our study opens an avenue to explore the p H-sensitive subcellular components as novel therapeutic targets for cancer treatment.
基金the Clinical Research Ethics Committee of the People’s Hospital of Guangxi Zhuang Autonomous Region(No.2013-09)。
文摘Background and Aims:Intrahepatic cholangiocarcinoma(ICC)is a malignant tumor derived from intrahepatic bile duct epithelial cells.Accumulating studies report that microRNAs are widely involved in tumor migration and metastasis by regulation of target genes.miR-7-5p has been confirmed to inhibit tumor metastasis and to be related to prognosis for several malignant tumors.Our study investigated the underlying functions of miR-7-5p in ICC.Methods:The expression of miR-7-5p in ICC tissues but also in ICC cell lines was analyzed by real-time PCR.By analyzing the relationship between the clinicopathological parameters of 60 ICC patients and the expression level of miR-7-5p,the effect of miR-7-5p on the prognosis was clarified.After transfected with miR-7-5p mimics or miR-7-5p inhibitor,cell counting kit-8 assay was applied to evaluate the cells proliferation,flow cytometry was applied to analyze the cells apoptosis,wound healing assay and transwell chamber assay were applied to analyze the cell invasion and migration.A luciferase reporter assay was identified the relationship of miR-7-5p and myeloid differentiation factor 88(MyD88).Western blotting was used to analyze the proteins expression.And immunochemistry was performed to determine the expression of MYD88 in ICC tissues.Results:Our data showed the expression of miR-7-5p was down-regulated not only in ICC tissues but also in ICC cell lines compared with normal controls.Low expression of miR-7-5p was notably associated with poor prognosis in ICC patients.miR-7-5p negatively regulated cell proliferation,migration,invasion and apoptosis in ICC cells.We further verified that MyD88 was a novel target of miR-7-5p and was significantly overexpressed in ICC tissues.Overexpression of MyD88 counteracted the effects of miR-7-5p in ICC cells.Conclusions:The present findings suggest that miR-7-5p plays a pivotal role in ICC invasion by regulating MyD88.Ampliative insight into the key factors of ICC invasion may result in the development of new treatment options for ICC.
