LncRNA PURPL调节miR-342-3p/PIK3R1轴对肝癌细胞恶性生物学行为的影响

doi:10.3969/j.issn.1006-6233.2024.01.05

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摘要 目的: 探究长链非编码RNA(PURPL)调节微小RNA-342-3p(miR-342-3p)/磷脂酰肌醇-3激酶调节亚基1(PIK3R1)轴对肝癌细胞恶性生物学行为的影响。方法: 细胞实验:将si-NC、si-PURPL、si-PURPL+inhibitor NC、si-PURPL+miR-342-3p inhibitor分别转染至肝癌细胞Huh-7细胞并命名为si-NC组、si-PURPL组、si-PURPL+inhibitor NC组、si-PURPL+miR-342-3p inhibitor 组,未做任何处理的Huh-7细胞记为NC组。双荧光素酶报告基因实验验证LncRNA PURPL、miR-342-3p、PIK3R1的关系;qRT-PCR检测人正常肝细胞系L02和人肝癌细胞系Huh-7中LncRNA PURPL、miR-342-3p 表达;Western blot检测L02细胞、Huh-7细胞PIK3R1蛋白水平;MTT法检测Huh-7细胞增殖情况;流式细胞术检测Huh-7细胞凋亡率;Transwell检测Huh-7细胞侵袭数量;划痕试验测定Huh-7细胞迁移。体内实验:构建裸鼠异种移植模型,分组同细胞实验,检测肿瘤体积和肿瘤重量。结果: 细胞实验结果显示:与si-NC组相比,si-PURPL组Huh-7细胞OD490值、划痕愈合率、侵袭细胞数量、LncRNA PURPL、PIK3R1水平显著下降(P<0.05),Huh-7细胞凋亡率、miR-342-3p相对表达量显著升高(P<0.05),而抑制miR-342-3p表达减弱了沉默LncRNA PURPL抑制Huh-7细胞增殖、迁移、侵袭和促进凋亡的效果;LncRNA PURPL负向调控miR-342-3p/PIK3R1轴。体内结果显示:si-PURPL组裸鼠肿瘤体积和质量较si-NC组下降(P<0.05);抑制miR-342-3p表达减弱了沉默LncRNA PURPL对体内肿瘤生长的抑制作用。结论: 沉默LncRNA PURPL可抑制Huh-7细胞的恶性生物学行为,其机制可能与调控miR-342-3p/PIK3R1轴有关。

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	王红梅
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关键词 ： LncRNA PURPL, miR-342-3p/PIK3R1轴, 肝癌, 增殖, 凋亡

Abstract：Objective: To investigate the impact of long non-coding RNA (PURPL) on the miR-342-3p/phosphoinositide-3-kinase regulatory subunit 1 (PIK3R1) axis and its effects on the malignant biological behavior of hepatocellular carcinoma (HCC) cells. Methods: Cell Experiments: Si-NC, si-PURPL, si-PURPL+inhibitor NC, si-PURPL+miR-342-3p inhibitor were separately transfected into HCC cells (Huh-7) and named si-NC group, si-PURPL group, si-PURPL+inhibitor NC group, si-PURPL+miR-342-3p inhibitor group, while untreated Huh-7 cells were designated as the NC group. Dual-luciferase reporter gene experiments verified the relationship between LncRNA PURPL, miR-342-3p, and PIK3R1. qRT-PCR was used to detect the expression of LncRNA PURPL and miR-342-3p in human normal liver cells (L02) and Huh-7 cells. Western blot assessed PIK3R1 protein levels in L02 and Huh-7 cells. MTT assay evaluated Huh-7 cell proliferation, flow cytometry measured apoptosis rates, Transwell assay determined invasive cell numbers, and scratch tests assessed cell migration. In Vivo Experiments: A xenograft mouse model was established with the same cell treatment groups. Tumor volume and weight were measured. Results: Cell experiment results showed that compared with the si-NC group, si-PURPL group had significantly decreased OD490 values, scratch healing rates, invasive cell numbers, LncRNA PURPL, and PIK3R1 levels in Huh-7 cells (P<0.05). Apoptosis rates and relative expression of miR-342-3p significantly increased (P<0.05). Inhibiting miR-342-3p expression weakened the inhibitory effects of silencing LncRNA PURPL on Huh-7 cell proliferation, migration, invasion, and promotion of apoptosis. LncRNA PURPL negatively regulated the miR-342-3p/PIK3R1 axis. In vivo results showed that the tumor volume and mass of si-PURPL group mice decreased compared to the si-NC group (P<0.05). Inhibiting miR-342-3p expression weakened the inhibitory effect of silencing LncRNA PURPL on tumor growth in vivo. Conclusion: Silting LncRNA PURPL can inhibit the malignant biological behavior of Huh-7 cells, and the mechanism may be related to the regulation of miR-342-3p/PIK3R1 axis.

Key words： LncRNA PURPL miR-342-3p/PIK3R1 axis Liver cancer Proliferation Apoptosis

基金资助:2019年四川省卫生健康委员会科研课题(重点研究项目),(编号:19ZD003)

引用本文:

王红梅, 陈思瑞, 杜红. LncRNA PURPL调节miR-342-3p/PIK3R1轴对肝癌细胞恶性生物学行为的影响[J]. 河北医学, 2024, 30(1): 29-35.
WANG Hongmei, CHEN Sirui, DU Hong. The Regulatory Effect of LncRNA PURPL on the miR-342-3p/PIK3R1 Axis and its Impact on the Malignant Biological Behavior of Hepatocellular Carcinoma Cells. HeBei Med, 2024, 30(1): 29-35.

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http://www.hbyxzzs.cn/CN/10.3969/j.issn.1006-6233.2024.01.05 或 http://www.hbyxzzs.cn/CN/Y2024/V30/I1/29

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