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

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

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。

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 experi-ment 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).In-hibiting 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 de-creased 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.