Objective This study aimed to examine the role of long non-coding RNA PCED1B antisense RNA 1(PCED1B-AS1)in the development of hepatocellular carcinoma(HCC).Methods A total of 62 pairs of HCC tissues and adjacent non-t...Objective This study aimed to examine the role of long non-coding RNA PCED1B antisense RNA 1(PCED1B-AS1)in the development of hepatocellular carcinoma(HCC).Methods A total of 62 pairs of HCC tissues and adjacent non-tumor tissues were obtained from 62 HCC patients.The interactions of PCED1B-AS1 and microRNA-34a(miR-34a)were detected by dual luciferase activity assay and RNA pull-down assay.The RNA expression levels of PCED1B-AS1,miR-34a and CD44 were detected by RT-qPCR,and the protein expression level of CD44 was determined by Western blotting.The cell proliferation was detected by cell proliferation assay,and the cell invasion and migration by transwell invasion assay.The HCC tumor growth after PCED1B-AS1 was downregulated was determined by in vivo animal study.Results PCED1B-AS1 was highly expressed in HCC tissues,which was associated with poor survival of HCC patients.Furthermore,PCED1B-AS1 interacted with miR-34a in HCC cells,but they did not regulate the expression of each other.Additionally,PCED1B-AS1 increased the expression level of CD44,which was targeted by miR-34a.The cell proliferation and invasion assay revealed that miR-34a inhibited the proliferation and invasion of HCC in vitro,while CD44 exhibited the opposite effects.Furthermore,PCED1B-AS1 suppressed the role of miR-34a.Moreover,the knockdown of PCED1B-AS1 repressed the HCC tumor growth in nude mice in vivo.Conclusion PCED1B-AS1 may play an oncogenic role by regulating the miR-34a/CD44 axis in HCC.展开更多
[目的]探讨长链非编码RNA(long non-coding RNA,lncRNA)PCED1B-AS1靶向miR-383-5p在结直肠癌(colorectal cancer,CRC)进展中的作用。[方法]收集解放军总医院第五医学中心南院区2018年1月至2019年10月收治的41例CRC组织和癌旁组织,RT-qPC...[目的]探讨长链非编码RNA(long non-coding RNA,lncRNA)PCED1B-AS1靶向miR-383-5p在结直肠癌(colorectal cancer,CRC)进展中的作用。[方法]收集解放军总医院第五医学中心南院区2018年1月至2019年10月收治的41例CRC组织和癌旁组织,RT-qPCR检测PCED1B-AS1、miR-383-5p和过氧化物酶3(PRDX3)mRNA表达水平。将si-NC、si-PCED1B-AS1、miR-NC、miR-383-5p、si-PCED1B-AS1+antimiR-NC、si-PCED1B-AS1+anti-miR-383-5p分别转染CRC细胞LoVo,CCK-8法、平板克隆实验、Transwell实验、流式细胞术分别检测LoVo细胞体外增殖、克隆形成、迁移和凋亡能力。双荧光素酶报告实验确定PCED1B-AS1与miR-383-5p、miR-383-5p与PRDX3的靶向关系。[结果]与癌旁组织比较,CRC组织中PCED1B-AS1(0.92±0.12 vs 2.87±0.32)、PRDX3 mRNA(0.93±0.11 vs 2.30±0.26)表达水平升高(P均<0.001),miR-383-5p表达水平降低(0.98±0.15 vs 0.40±0.05,P<0.001)。下调PCED1B-AS1后LoVo细胞miR-383-5p表达水平(1.00±0.00 vs 2.95±0.12)、抑制率(0 vs 47.75%±2.75%)、凋亡率均升高(8.10%±0.75%vs 20.20%±1.09%)(P均<0.001),PRDX3 mRNA表达水平(1.00±0.00 vs 0.27±0.03)、克隆形成数(116.78±6.01 vs67.56±4.11)、迁移细胞数(173.78±8.32 vs 85.89±3.07)均降低(P均<0.001)。过表达miR-383-5p后LoVo细胞PRDX3 m RNA表达水平、克隆形成数、迁移细胞数降低(P均<0.001),抑制率、凋亡率升高(P均<0.001)。miR-383-5p分别与PCED1B-AS1、PRDX3特异性结合。抑制miR-383-5p表达可减弱下调PCED1B-AS1对LoVo细胞增殖、迁移、克隆形成、凋亡的影响(P均<0.001)。[结论]下调PCED1B-AS1通过靶向上调miR-383-5p表达可抑制CRC细胞增殖和迁移,诱导细胞凋亡,进而抑制CRC进展。展开更多
基金supported by the Medical Science and Technology Research Foundation of Guangdong Province(No.A2020559).
文摘Objective This study aimed to examine the role of long non-coding RNA PCED1B antisense RNA 1(PCED1B-AS1)in the development of hepatocellular carcinoma(HCC).Methods A total of 62 pairs of HCC tissues and adjacent non-tumor tissues were obtained from 62 HCC patients.The interactions of PCED1B-AS1 and microRNA-34a(miR-34a)were detected by dual luciferase activity assay and RNA pull-down assay.The RNA expression levels of PCED1B-AS1,miR-34a and CD44 were detected by RT-qPCR,and the protein expression level of CD44 was determined by Western blotting.The cell proliferation was detected by cell proliferation assay,and the cell invasion and migration by transwell invasion assay.The HCC tumor growth after PCED1B-AS1 was downregulated was determined by in vivo animal study.Results PCED1B-AS1 was highly expressed in HCC tissues,which was associated with poor survival of HCC patients.Furthermore,PCED1B-AS1 interacted with miR-34a in HCC cells,but they did not regulate the expression of each other.Additionally,PCED1B-AS1 increased the expression level of CD44,which was targeted by miR-34a.The cell proliferation and invasion assay revealed that miR-34a inhibited the proliferation and invasion of HCC in vitro,while CD44 exhibited the opposite effects.Furthermore,PCED1B-AS1 suppressed the role of miR-34a.Moreover,the knockdown of PCED1B-AS1 repressed the HCC tumor growth in nude mice in vivo.Conclusion PCED1B-AS1 may play an oncogenic role by regulating the miR-34a/CD44 axis in HCC.
文摘[目的]探讨长链非编码RNA(long non-coding RNA,lncRNA)PCED1B-AS1靶向miR-383-5p在结直肠癌(colorectal cancer,CRC)进展中的作用。[方法]收集解放军总医院第五医学中心南院区2018年1月至2019年10月收治的41例CRC组织和癌旁组织,RT-qPCR检测PCED1B-AS1、miR-383-5p和过氧化物酶3(PRDX3)mRNA表达水平。将si-NC、si-PCED1B-AS1、miR-NC、miR-383-5p、si-PCED1B-AS1+antimiR-NC、si-PCED1B-AS1+anti-miR-383-5p分别转染CRC细胞LoVo,CCK-8法、平板克隆实验、Transwell实验、流式细胞术分别检测LoVo细胞体外增殖、克隆形成、迁移和凋亡能力。双荧光素酶报告实验确定PCED1B-AS1与miR-383-5p、miR-383-5p与PRDX3的靶向关系。[结果]与癌旁组织比较,CRC组织中PCED1B-AS1(0.92±0.12 vs 2.87±0.32)、PRDX3 mRNA(0.93±0.11 vs 2.30±0.26)表达水平升高(P均<0.001),miR-383-5p表达水平降低(0.98±0.15 vs 0.40±0.05,P<0.001)。下调PCED1B-AS1后LoVo细胞miR-383-5p表达水平(1.00±0.00 vs 2.95±0.12)、抑制率(0 vs 47.75%±2.75%)、凋亡率均升高(8.10%±0.75%vs 20.20%±1.09%)(P均<0.001),PRDX3 mRNA表达水平(1.00±0.00 vs 0.27±0.03)、克隆形成数(116.78±6.01 vs67.56±4.11)、迁移细胞数(173.78±8.32 vs 85.89±3.07)均降低(P均<0.001)。过表达miR-383-5p后LoVo细胞PRDX3 m RNA表达水平、克隆形成数、迁移细胞数降低(P均<0.001),抑制率、凋亡率升高(P均<0.001)。miR-383-5p分别与PCED1B-AS1、PRDX3特异性结合。抑制miR-383-5p表达可减弱下调PCED1B-AS1对LoVo细胞增殖、迁移、克隆形成、凋亡的影响(P均<0.001)。[结论]下调PCED1B-AS1通过靶向上调miR-383-5p表达可抑制CRC细胞增殖和迁移,诱导细胞凋亡,进而抑制CRC进展。
