| Zunli Yi,Xiaoguang Guo,Xianxue Jiang,Fengmei Luo. Effects of long non-coding RNA GAS5 on proliferation and apoptosis of hepatocellular carcinoma cells through miR-26a-5p action. Oncol Transl Med, 2022, 8: 126-134. |
| 长链非编码RNA GAS5通过miR-26a-5p对肝癌细胞增殖和凋亡的影响 |
| Effects of long non-coding RNA GAS5 on proliferation and apoptosis of hepatocellular carcinoma cells through miR-26a-5p action |
| Received:July 07, 2021 Revised:June 29, 2022 |
| DOI：10.1007/s10330-021-0507-7 |
| 中文关键词: lncRNA GAS5;miR-26a-5p;肝细胞carcinomar;细胞增殖;细胞周期;细胞凋亡 |
| 英文关键词: lncRNA GAS5; miR-26a-5p; hepatocellular carcinoma (HCC); cell proliferation; cell cycle; apoptosis |
| 基金项目: |
| Author Name | Affiliation | E-mail | | Zunli Yi* | Pathology Department, Yuechi People''s Hospital | zhuichengkong2351@163.com | | Xiaoguang Guo | Pathology Department, Nanchong Central Hospital | | | Xianxue Jiang | The First Department of General Surgery, Yuechi People''s Hospital | | | Fengmei Luo | The Second Department of General Surgery, Yuechi People''s Hospital | |
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| 中文摘要: |
| 摘要目的:探讨长链非编码RNA(lncRNA)生长阻滞特殊5(GAS5)对肝细胞癌(HCC)细胞生物学行为的影响及其机制。方法:采用RT-qPCR检测80例HCC患者癌组织和癌旁组织中lncRNA GAS5和miR-26a-5p的表达水平。Starbase网站预测和荧光素酶基因报告分析lncRNA GAS5与miR-26a-5p的关系。将质粒转染人肝癌细胞系SMMC-7721细胞，分为NC组、lncRNA GAS5组、anti-miR-26a-5p组、lncRNA GAS5+miR-26a-5p组。检测各组细胞增殖、细胞周期及凋亡情况。通过TargetScan数据库预测和荧光素酶基因报告分析miR-26a-5p和PTEN之间的关系。Western Blot检测磷酸酶，确定染色体磷脂酰肌醇3-激酶PI3K、磷酸化蛋白激酶B (p-Akt)、Cyclin D1(Cycle)蛋白在Cyclin D1、Cyclin D1和人P27蛋白(P27)中表达。结果:(1)与癌旁组织相比，lncRNA GAS5在HCC组织中的表达明显降低(P<0.05);miR-26a-5p的表达明显增加(P < 0.05)。(2)Starbase预测和荧光素酶基因的报道结果表明miR-26a-5p有针对性的结合位点与lncRNA GAS5。(3)与NC组相比,EDU阳性细胞率在lncRNA GAS5组和anti-miR-26a-5p组显著降低,G0/G1期细胞数量显著增加，s期细胞数量显著减少，差异均有统计学意义(P<0.05)。与lncRNA GAS5组相比，lncRNA GAS5+miR-26a-5p组EDU阳性细胞率显著增加，G0/G1期细胞数显著减少，S期细胞数显著增加，(5)与NC组相比，lncRNA GAS5组和anti-miR-26a-5p组中PTEN和P27表达增加，而PI3K、miR-26a-5p、miR-26a-5p、miR-26a-5p、miR-26a-5p、miR-26a-5p、miR-26a-5p、miR-26a-5p、miR-26a-5p、miR-26a-5p、P - akt和Cyclin D1降低，差异均有统计学意义(P<0.05)。与lncRNA GAS5组相比，lncRNA GAS5+miR-26a-5p组PTEN、P27表达降低，PI3K、P - akt、Cyclin D1表达升高(P<0.05)。结论:lncRNA GAS5在HCC中表达降低，并通过miR-26a-5p调控PTNE/PI3K/Akt信号通路调控肝癌细胞增殖和凋亡。 |
| 英文摘要: |
| Objective Long non-coding RNAs (lncRNAs) regulate tumor development and progression by promoting
tumor proliferation, invasion, and metastasis. The aim of the study was to investigate the effects of lncRNA
growth arrest-special 5 (GAS5) on proliferation and apoptosis of hepatocellular carcinoma (HCC) cells
through miR-26a-5p action.
Methods Expression levels of GAS5 were detected in cancerous and paracancerous tissue of 80 HCC
patients by RT-qPCR. The starBase tool predicted that GAS5 had binding sites for the miRNA miR-26a-5p,
which was also highly expressed in HCC tissue. The relationship between GAS5 and miR-26a-5p was
confirmed using a luciferase reporter assay. The role of these lncRNAs was further explored by transfecting
plasmids into SMMC-7721 cells and classifying the cells as follows: NC group, GAS5 group, anti-miR-
26a-5p group, and GAS5 + miR-26a-5p group. Cell proliferation, cell cycle, and apoptosis were detected
in each group. The relationship between miR-26a-5p and phosphatase and tensin homolog deleted on
chromosome 10 (PTEN) was analyzed by TargetScan database prediction and luciferase reporter assay.
Western blotting was used to quantify PTEN, phosphatidylinositol 3-kinase (PI3K), phosphorylated protein
kinase B (p-Akt), cyclin D1, and human P27 protein (P27).
Results GAS5 was downregulated, while miR-26a-5p was upregulated in HCC tissue compared to in
paracancerous tissue. High GAS5 levels and low miR-26a-5p levels inhibited cell proliferation, increased
the number of G0/G1 phase cells, promoted cell apoptosis, promoted PTEN and P27 expression, and
inhibited PI3K, P-Akt, and cyclin D1 expression at the protein level. Upregulation of miR-26a-5p attenuated
the effects of GAS5 upregulation on the proliferation, cell cycle, and apoptosis of HCC cells and on the
expression of PTNE/PI3K/Akt signaling pathway-related proteins.
Conclusion Low GAS5 levels regulate the proliferation and apoptosis of HCC cells via the PTNE/PI3K/
Akt signaling pathway and are linked to upregulation of miR-26a-5p. |
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