Hepatocellular carcinoma(HCC)is a malignancy known for its unfavorable prognosis.The dysregulation of the tumor microenvironment(TME)can affect the sensitivity to immunotherapy or chemotherapy,leading to treatment fai...Hepatocellular carcinoma(HCC)is a malignancy known for its unfavorable prognosis.The dysregulation of the tumor microenvironment(TME)can affect the sensitivity to immunotherapy or chemotherapy,leading to treatment failure.The elucidation of PHLDA2’s involvement in HCC is imperative,and the clinical value of PHLDA2 is also underestimated.Here,bioinformatics analysis was performed in multiple cohorts to explore the phenotype and mechanism through which PHLDA2 may affect the progression of HCC.Then,the expression and function of PHLDA2 were examined via the qRT-PCR,Western Blot,and MTT assays.Our findings indicate a substantial upregulation of PHLDA2 in HCC,correlated with a poorer prognosis.The methylation levels of PHLDA2 were found to be lower in HCC tissues compared to normal liver tissues.Besides,noteworthy associations were observed between PHLDA2 expression and immune infiltration in HCC.In addition,PHLDA2 upregulation is closely associated with stemness features and immunotherapy or chemotherapy resistance in HCC.In vitro experiments showed that sorafenib or cisplatin significantly up-regulated PHLDA2 mRNA levels,and PHLDA2 knockdown markedly decreased the sensitivity of HCC cells to chemotherapy drugs.Meanwhile,we found that TGF-βinduced the expression of PHLDA2 in vitro.The GSEA and in vitro experiment indicated that PHLDA2 may promote the HCC progression via activating the AKT signaling pathway.Our study revealed the novel role of PHLDA2 as an independent prognostic factor,which plays an essential role in TME remodeling and treatment resistance in HCC.展开更多
为了研究山羊血小板白细胞C激酶底物同源性样结构域蛋白家族A成员2 (pleckstrin homology-like domain family A member 2, PHLDA2)基因表达水平与其启动子甲基化状态相关性,试验采用RT-PCR方法检测PHLDA2在各组织间的表达水平(心脏、...为了研究山羊血小板白细胞C激酶底物同源性样结构域蛋白家族A成员2 (pleckstrin homology-like domain family A member 2, PHLDA2)基因表达水平与其启动子甲基化状态相关性,试验采用RT-PCR方法检测PHLDA2在各组织间的表达水平(心脏、肝脏、脾脏、肺脏、肾脏、肌肉、脂肪、舌、大脑、胎盘),以及扩增了山羊PHLDA2基因启动子区域序列,并利用Methylation specific PCR技术检测CpG岛在山羊各组织间的甲基化水平。结果表明:克隆得到的山羊PHLDA2 1842 bp的启动子序列与绵羊、牛PHLDA2启动子同源性分别达93.2%和87.9%;启动区CpG岛内第三位点CpG的甲基化频率和胎盘第一外显子区域甲基化水平显著低于其他组织(P 【0.05)。说明PHLDA2的mRNA相对表达量水平与启动子区CpG岛(−302~−88 bp)内第三个CpG位点甲基化频率和第一外显子CpG岛(49~333 bp)甲基化水平呈负相关。展开更多
基金supported by the National Natural Science Foundation of China(Nos.81872255,62141109)the Leading-Edge Technology Programme of Jiangsu Natural Science Foundation:BK20212021.
文摘Hepatocellular carcinoma(HCC)is a malignancy known for its unfavorable prognosis.The dysregulation of the tumor microenvironment(TME)can affect the sensitivity to immunotherapy or chemotherapy,leading to treatment failure.The elucidation of PHLDA2’s involvement in HCC is imperative,and the clinical value of PHLDA2 is also underestimated.Here,bioinformatics analysis was performed in multiple cohorts to explore the phenotype and mechanism through which PHLDA2 may affect the progression of HCC.Then,the expression and function of PHLDA2 were examined via the qRT-PCR,Western Blot,and MTT assays.Our findings indicate a substantial upregulation of PHLDA2 in HCC,correlated with a poorer prognosis.The methylation levels of PHLDA2 were found to be lower in HCC tissues compared to normal liver tissues.Besides,noteworthy associations were observed between PHLDA2 expression and immune infiltration in HCC.In addition,PHLDA2 upregulation is closely associated with stemness features and immunotherapy or chemotherapy resistance in HCC.In vitro experiments showed that sorafenib or cisplatin significantly up-regulated PHLDA2 mRNA levels,and PHLDA2 knockdown markedly decreased the sensitivity of HCC cells to chemotherapy drugs.Meanwhile,we found that TGF-βinduced the expression of PHLDA2 in vitro.The GSEA and in vitro experiment indicated that PHLDA2 may promote the HCC progression via activating the AKT signaling pathway.Our study revealed the novel role of PHLDA2 as an independent prognostic factor,which plays an essential role in TME remodeling and treatment resistance in HCC.
文摘为了研究山羊血小板白细胞C激酶底物同源性样结构域蛋白家族A成员2 (pleckstrin homology-like domain family A member 2, PHLDA2)基因表达水平与其启动子甲基化状态相关性,试验采用RT-PCR方法检测PHLDA2在各组织间的表达水平(心脏、肝脏、脾脏、肺脏、肾脏、肌肉、脂肪、舌、大脑、胎盘),以及扩增了山羊PHLDA2基因启动子区域序列,并利用Methylation specific PCR技术检测CpG岛在山羊各组织间的甲基化水平。结果表明:克隆得到的山羊PHLDA2 1842 bp的启动子序列与绵羊、牛PHLDA2启动子同源性分别达93.2%和87.9%;启动区CpG岛内第三位点CpG的甲基化频率和胎盘第一外显子区域甲基化水平显著低于其他组织(P 【0.05)。说明PHLDA2的mRNA相对表达量水平与启动子区CpG岛(−302~−88 bp)内第三个CpG位点甲基化频率和第一外显子CpG岛(49~333 bp)甲基化水平呈负相关。
