Objective Resveratrol(Res)is a promising anticancer drug against hepatocellular carcinoma(HCC),but whether its anti-HCC effects implicate mitophagy remains unclear.Therefore,we aimed to explore the specific role of Re...Objective Resveratrol(Res)is a promising anticancer drug against hepatocellular carcinoma(HCC),but whether its anti-HCC effects implicate mitophagy remains unclear.Therefore,we aimed to explore the specific role of Res in mitophagy and the related mechanisms during the treatment of HCC.Methods HepG2 cells and tumor-grafted nude mice were used to investigate the effects of low-,middle-and high-dose of Res on HCC progression and mitophagy in vitro and in vivo,respectively.A series of approaches including cell counting kit-8,flow cytometry,wound healing and transwell assays were used to evaluate tumor cell functions.Transmission electron microscopy,immunofluorescence and Western blotting analysis were used to assess mitophagy.Mitochondrial oxygen consumption rate,reactive oxygen species and membrane potential were used to reflect mitochondrial function.After disrupting the expression of metastasis-associated lung adenocarcinoma transcript 1(MALAT1),miR-143-3p,and ribonucleoside reductase M2(RRM2),the effects of the MALAT1/miR-143-3p/RRM2 axis on cell function and mitophagy under Res treatment were explored in vitro.Additionally,dual-luciferase reporter and chromatin immunoprecipitation were used to confirm interactions between target genes.Results Res significantly inhibited the proliferation and promoted apoptosis of HCC cells in vitro,while significantly suppressing tumor growth in a dose-dependent manner and inducing mitophagy and mitochondrial dysfunction in vivo.Interestingly,MALAT1 was highly expressed in HCC cells and its knockdown upregulated miR-143-3p expression in HCC cells,which subsequently inhibited RRM2 expression.Furthermore,in nude mice grafted with HCC tumors and treated with Res,the expression of MALAT1,miR-143-3p and RRM2 were altered significantly.In vitro data further supported the targeted binding relationships between MALAT1 and miR-143-3p and between miR-143-3p and RRM2.Therefore,a series of cell-based experiments were carried out to study the mechanism of the MALAT1/miR-143-3p/RRM2 axis involved in mitophagy and HCC;these experiments revealed that MALAT1 knockdown,miR-143-3p mimic and RRM silencing potentiated the antitumor effects of Res and its activation of mitophagy.Conclusion Res facilitated mitophagy in HCC and exerted anti-cancer effects by targeting the MALAT1/miR-143-3p/RRM2 axis.展开更多
目的探讨颅内动脉瘤患者血清缺氧诱导因子-1α(hypoxia-inducible factor-1α,HIF-1α)、半胱氨酸天冬氨酸蛋白水解酶-3(cysteine aspartate protein hydrolase-3,Caspase-3)、微小RNA-143(microRNA-143,miR-143)水平与动脉瘤破裂的关...目的探讨颅内动脉瘤患者血清缺氧诱导因子-1α(hypoxia-inducible factor-1α,HIF-1α)、半胱氨酸天冬氨酸蛋白水解酶-3(cysteine aspartate protein hydrolase-3,Caspase-3)、微小RNA-143(microRNA-143,miR-143)水平与动脉瘤破裂的关系。方法143例颅内动脉瘤患者作为病例组,根据颅内动脉瘤破裂与否分为未破裂组(n=95)和破裂组(n=48),另选取同期健康体检人群138例作为对照组,收集各组受检者的一般资料(年龄、性别及体质量指标);抽取病例组患者入院日、对照组体检者体检日空腹肘静脉血离心,采用酶联免疫吸附试验和实时荧光定量PCR检测各组受检者血清HIF-1α、Caspase-3及miR-143水平;随访病例组患者术后3个月,采用CT血管造影检查患者的动脉瘤影像特征(动脉瘤标准化表面最大压力、瘤体宽径、形状不规则占比、载瘤动脉宽度、低剪切力面积比值、动脉瘤标准化表面平均压力、瘤体长径、瘤径宽度、瘤体最长径、平均壁面剪切力及动脉瘤位置)并判断颅内动脉瘤破裂与否,采用多因素logistic分析病例组患者颅内动脉瘤破裂的影响因素,采用Spearman相关分析和ROC曲线分析病例组患者血清HIF-1α、Caspase-3、miR-143水平与颅内动脉瘤破裂的相关性及预警价值。结果病例组患者血清Caspase-3、HIF-1α高于对照组,miR-143水平低于对照组(P<0.05);破裂组颅内动脉瘤患者血清miR-143水平和瘤体宽径低于未破裂组,血清HIF-1α、Caspase-3及低剪切力面积比值、瘤体最长径、形状不规则占比、瘤体长径、平均壁面剪切力水平高于未破裂组(P<0.05);logistic分析结果显示,血清Caspase-3、miR-143、HIF-1α及低剪切力面积比值、瘤体宽径、瘤体长径、平均壁面剪切力、瘤体最长径、形状不规则是颅内动脉瘤破裂的影响因素(P<0.05);Spearman相关分析结果显示,血清miR-143与患者颅内动脉瘤破裂呈负相关(P<0.05),血清Caspase-3、HIF-1α与患者颅内动脉瘤破裂呈正相关(P<0.05);血清miR-143、Caspase-3及HIF-1α预测颅内动脉瘤破裂的曲线下面积(AUC)分别为0.716、0.689及0.702,3者联合预测颅内动脉瘤破裂的AUC为0.802大于单独指标预测。结论血清高HIF-1α、Caspase-3水平及低miR-143水平是颅内动脉瘤破裂的影响因素,且3者联合预测价值更高。展开更多
基金supported by the Zhejiang Provincial Science and Technology Department Key Research and Development Plan(No.2020C03046).
文摘Objective Resveratrol(Res)is a promising anticancer drug against hepatocellular carcinoma(HCC),but whether its anti-HCC effects implicate mitophagy remains unclear.Therefore,we aimed to explore the specific role of Res in mitophagy and the related mechanisms during the treatment of HCC.Methods HepG2 cells and tumor-grafted nude mice were used to investigate the effects of low-,middle-and high-dose of Res on HCC progression and mitophagy in vitro and in vivo,respectively.A series of approaches including cell counting kit-8,flow cytometry,wound healing and transwell assays were used to evaluate tumor cell functions.Transmission electron microscopy,immunofluorescence and Western blotting analysis were used to assess mitophagy.Mitochondrial oxygen consumption rate,reactive oxygen species and membrane potential were used to reflect mitochondrial function.After disrupting the expression of metastasis-associated lung adenocarcinoma transcript 1(MALAT1),miR-143-3p,and ribonucleoside reductase M2(RRM2),the effects of the MALAT1/miR-143-3p/RRM2 axis on cell function and mitophagy under Res treatment were explored in vitro.Additionally,dual-luciferase reporter and chromatin immunoprecipitation were used to confirm interactions between target genes.Results Res significantly inhibited the proliferation and promoted apoptosis of HCC cells in vitro,while significantly suppressing tumor growth in a dose-dependent manner and inducing mitophagy and mitochondrial dysfunction in vivo.Interestingly,MALAT1 was highly expressed in HCC cells and its knockdown upregulated miR-143-3p expression in HCC cells,which subsequently inhibited RRM2 expression.Furthermore,in nude mice grafted with HCC tumors and treated with Res,the expression of MALAT1,miR-143-3p and RRM2 were altered significantly.In vitro data further supported the targeted binding relationships between MALAT1 and miR-143-3p and between miR-143-3p and RRM2.Therefore,a series of cell-based experiments were carried out to study the mechanism of the MALAT1/miR-143-3p/RRM2 axis involved in mitophagy and HCC;these experiments revealed that MALAT1 knockdown,miR-143-3p mimic and RRM silencing potentiated the antitumor effects of Res and its activation of mitophagy.Conclusion Res facilitated mitophagy in HCC and exerted anti-cancer effects by targeting the MALAT1/miR-143-3p/RRM2 axis.
文摘目的探讨颅内动脉瘤患者血清缺氧诱导因子-1α(hypoxia-inducible factor-1α,HIF-1α)、半胱氨酸天冬氨酸蛋白水解酶-3(cysteine aspartate protein hydrolase-3,Caspase-3)、微小RNA-143(microRNA-143,miR-143)水平与动脉瘤破裂的关系。方法143例颅内动脉瘤患者作为病例组,根据颅内动脉瘤破裂与否分为未破裂组(n=95)和破裂组(n=48),另选取同期健康体检人群138例作为对照组,收集各组受检者的一般资料(年龄、性别及体质量指标);抽取病例组患者入院日、对照组体检者体检日空腹肘静脉血离心,采用酶联免疫吸附试验和实时荧光定量PCR检测各组受检者血清HIF-1α、Caspase-3及miR-143水平;随访病例组患者术后3个月,采用CT血管造影检查患者的动脉瘤影像特征(动脉瘤标准化表面最大压力、瘤体宽径、形状不规则占比、载瘤动脉宽度、低剪切力面积比值、动脉瘤标准化表面平均压力、瘤体长径、瘤径宽度、瘤体最长径、平均壁面剪切力及动脉瘤位置)并判断颅内动脉瘤破裂与否,采用多因素logistic分析病例组患者颅内动脉瘤破裂的影响因素,采用Spearman相关分析和ROC曲线分析病例组患者血清HIF-1α、Caspase-3、miR-143水平与颅内动脉瘤破裂的相关性及预警价值。结果病例组患者血清Caspase-3、HIF-1α高于对照组,miR-143水平低于对照组(P<0.05);破裂组颅内动脉瘤患者血清miR-143水平和瘤体宽径低于未破裂组,血清HIF-1α、Caspase-3及低剪切力面积比值、瘤体最长径、形状不规则占比、瘤体长径、平均壁面剪切力水平高于未破裂组(P<0.05);logistic分析结果显示,血清Caspase-3、miR-143、HIF-1α及低剪切力面积比值、瘤体宽径、瘤体长径、平均壁面剪切力、瘤体最长径、形状不规则是颅内动脉瘤破裂的影响因素(P<0.05);Spearman相关分析结果显示,血清miR-143与患者颅内动脉瘤破裂呈负相关(P<0.05),血清Caspase-3、HIF-1α与患者颅内动脉瘤破裂呈正相关(P<0.05);血清miR-143、Caspase-3及HIF-1α预测颅内动脉瘤破裂的曲线下面积(AUC)分别为0.716、0.689及0.702,3者联合预测颅内动脉瘤破裂的AUC为0.802大于单独指标预测。结论血清高HIF-1α、Caspase-3水平及低miR-143水平是颅内动脉瘤破裂的影响因素,且3者联合预测价值更高。
