[目的]探究miR-552-3p调节SETD2对肾细胞癌786-O细胞转移活性的影响。[方法]通过实时的荧光定量PCR方式对正常肾小管上皮细胞株HK-2细胞、透明细胞肾细胞癌细胞株786-O细胞、RCC23细胞、Caki-2细胞中的miR-552-3p与SETD2表达情况进行检...[目的]探究miR-552-3p调节SETD2对肾细胞癌786-O细胞转移活性的影响。[方法]通过实时的荧光定量PCR方式对正常肾小管上皮细胞株HK-2细胞、透明细胞肾细胞癌细胞株786-O细胞、RCC23细胞、Caki-2细胞中的miR-552-3p与SETD2表达情况进行检测。将786-O细胞分为miR NC组、miR-552-3p inhibitor组、pcDNA3.1 NC组以及pcDNA3.1 SETD2组。通过CCK-8实验分析786-O细胞增殖能力;Transwell实验和划痕实验分析786-O细胞侵袭和迁移能力;荧光素酶报告基因实验分析786-O细胞中miR-552-3p与SETD2的靶向关系。[结果]与正常肾小管上皮细胞株相比,miR-552-3p在透明细胞肾细胞癌细胞株中高表达(0.22±0.03 vs 0.92±0.05 vs 0.87±0.06 vs 0.88±0.11),而SETD2在透明细胞肾细胞癌细胞株中低表达(0.93±0.05 vs 0.31±0.03 vs 0.32±0.07 vs 0.29±0.06)。与miR NC组比较,miR-552-3p inhibitor组的786-O细胞的增殖活性降低(2.21±0.08 vs 1.36±0.11)、侵袭和迁移能力降低(103.19±9.38 vs 52.09±5.76个;73.19%±6.35%vs 22.12%±5.21%)。与pcDNA3.1 NC组比较,pcDNA3.1 SETD2组的786-O细胞的增殖活性降低、侵袭和迁移能力降低(P<0.05)。荧光素酶活性测定显示,和miR NC组比较,在SETD2-WT组中转染miR-552-3p mimic后荧光素酶活性降低(P<0.05)。[结论]miR-552-3p在透明细胞肾细胞癌细胞中高表达,而SETD2低表达。miR-552-3p能够抑制透明细胞肾细胞癌细胞的体外增殖、迁移和侵袭,并且这一作用与miR-552-3p抑制SETD2的表达相关。展开更多
BACKGROUND Hepatocellular carcinoma(HCC)has been a pervasive malignancy throughout the world with elevated mortality.Efficient therapeutic targets are beneficial to treat and predict the disease.Currently,the exact mo...BACKGROUND Hepatocellular carcinoma(HCC)has been a pervasive malignancy throughout the world with elevated mortality.Efficient therapeutic targets are beneficial to treat and predict the disease.Currently,the exact molecular mechanisms leading to the progression of HCC are still unclear.Research has shown that the microRNA-142-3p level decreases in HCC,whereas bioinformatics analysis of the cancer genome atlas database shows the ASH1L expression increased among liver tumor tissues.In this paper,we will explore the effects and mechanisms of microRNA-142-3p and ASH1L affect the prognosis of HCC patients and HCC cell bioactivity,and the association between them.AIM To investigate the effects and mechanisms of microRNA-142-3p and ASH1L on the HCC cell bioactivity and prognosis of HCC patients.METHODS In this study,we grouped HCC patients according to their immunohistochemistry results of ASH1L with pathological tissues,and retrospectively analyzed the prognosis of HCC patients.Furthermore,explored the roles and mechanisms of microRNA-142-3p and ASH1L by cellular and animal experiments,which involved the following experimental methods:Immunohistochemical staining,western blot,quantitative real-time-polymerase chain reaction,flow cytometric analysis,tumor xenografts in nude mice,etc.The statistical methods involved in this study contained t-test,one-way analysis of variance,theχ^(2)test,the Kaplan-Meier approach and the log-rank test.RESULTS In this study,we found that HCC patients with high expression of ASH1L possess a more recurrence rate as well as a decreased overall survival rate.ASH1L promotes the tumorigenicity of HCC and microRNA-142-3p exhibits reduced expression in HCC tissues and interacts with ASH1L through targeting the ASH1L 3′untranslated region.Furthermore,microRNA-142-3p promotes apoptosis and inhibits proliferation,invasion,and migration of HCC cell lines in vitro via ASH1L.For the exploration mechanism,we found ASH1L may promote an immunosuppressive microenvironment in HCC and ASH1L affects the expression of the cell junction protein zonula occludens-1,which is potentially relevant to the immune system.CONCLUSION Loss function of microRNA-142-3p induces cancer progression and immune evasion through upregulation of ASH1L in HCC.Both microRNA-142-3p and ASH1L can feature as new biomarker for HCC in the future.展开更多
文摘[目的]探究miR-552-3p调节SETD2对肾细胞癌786-O细胞转移活性的影响。[方法]通过实时的荧光定量PCR方式对正常肾小管上皮细胞株HK-2细胞、透明细胞肾细胞癌细胞株786-O细胞、RCC23细胞、Caki-2细胞中的miR-552-3p与SETD2表达情况进行检测。将786-O细胞分为miR NC组、miR-552-3p inhibitor组、pcDNA3.1 NC组以及pcDNA3.1 SETD2组。通过CCK-8实验分析786-O细胞增殖能力;Transwell实验和划痕实验分析786-O细胞侵袭和迁移能力;荧光素酶报告基因实验分析786-O细胞中miR-552-3p与SETD2的靶向关系。[结果]与正常肾小管上皮细胞株相比,miR-552-3p在透明细胞肾细胞癌细胞株中高表达(0.22±0.03 vs 0.92±0.05 vs 0.87±0.06 vs 0.88±0.11),而SETD2在透明细胞肾细胞癌细胞株中低表达(0.93±0.05 vs 0.31±0.03 vs 0.32±0.07 vs 0.29±0.06)。与miR NC组比较,miR-552-3p inhibitor组的786-O细胞的增殖活性降低(2.21±0.08 vs 1.36±0.11)、侵袭和迁移能力降低(103.19±9.38 vs 52.09±5.76个;73.19%±6.35%vs 22.12%±5.21%)。与pcDNA3.1 NC组比较,pcDNA3.1 SETD2组的786-O细胞的增殖活性降低、侵袭和迁移能力降低(P<0.05)。荧光素酶活性测定显示,和miR NC组比较,在SETD2-WT组中转染miR-552-3p mimic后荧光素酶活性降低(P<0.05)。[结论]miR-552-3p在透明细胞肾细胞癌细胞中高表达,而SETD2低表达。miR-552-3p能够抑制透明细胞肾细胞癌细胞的体外增殖、迁移和侵袭,并且这一作用与miR-552-3p抑制SETD2的表达相关。
基金Supported by the Haihe Laboratory of Cell Ecosystem Innovation Fund,No.22HHXBJC00001the Key Discipline Special Project of Tianjin Municipal Health Commission,No.TJWJ2022XK016.
文摘BACKGROUND Hepatocellular carcinoma(HCC)has been a pervasive malignancy throughout the world with elevated mortality.Efficient therapeutic targets are beneficial to treat and predict the disease.Currently,the exact molecular mechanisms leading to the progression of HCC are still unclear.Research has shown that the microRNA-142-3p level decreases in HCC,whereas bioinformatics analysis of the cancer genome atlas database shows the ASH1L expression increased among liver tumor tissues.In this paper,we will explore the effects and mechanisms of microRNA-142-3p and ASH1L affect the prognosis of HCC patients and HCC cell bioactivity,and the association between them.AIM To investigate the effects and mechanisms of microRNA-142-3p and ASH1L on the HCC cell bioactivity and prognosis of HCC patients.METHODS In this study,we grouped HCC patients according to their immunohistochemistry results of ASH1L with pathological tissues,and retrospectively analyzed the prognosis of HCC patients.Furthermore,explored the roles and mechanisms of microRNA-142-3p and ASH1L by cellular and animal experiments,which involved the following experimental methods:Immunohistochemical staining,western blot,quantitative real-time-polymerase chain reaction,flow cytometric analysis,tumor xenografts in nude mice,etc.The statistical methods involved in this study contained t-test,one-way analysis of variance,theχ^(2)test,the Kaplan-Meier approach and the log-rank test.RESULTS In this study,we found that HCC patients with high expression of ASH1L possess a more recurrence rate as well as a decreased overall survival rate.ASH1L promotes the tumorigenicity of HCC and microRNA-142-3p exhibits reduced expression in HCC tissues and interacts with ASH1L through targeting the ASH1L 3′untranslated region.Furthermore,microRNA-142-3p promotes apoptosis and inhibits proliferation,invasion,and migration of HCC cell lines in vitro via ASH1L.For the exploration mechanism,we found ASH1L may promote an immunosuppressive microenvironment in HCC and ASH1L affects the expression of the cell junction protein zonula occludens-1,which is potentially relevant to the immune system.CONCLUSION Loss function of microRNA-142-3p induces cancer progression and immune evasion through upregulation of ASH1L in HCC.Both microRNA-142-3p and ASH1L can feature as new biomarker for HCC in the future.
