长链非编码RNA LINC01410对B细胞非霍奇金淋巴瘤细胞增殖和凋亡的影响

Effects of long noncoding RNA LINC01410 on proliferation and apoptosis in B-cell non-Hodgkin′s lymphoma cells

目的探讨长链非编码RNA(LncRNA)LINC01410对B细胞非霍奇金淋巴瘤(B-NHL)细胞增殖和凋亡的影响。方法采用Cox回归模型筛选GEO数据集(GSE10846)中与B-NHL预后相关的LncRNA。将体外培养B-NHL细胞系Raji和Ramos,随机分为慢病毒对照组、LINC01410敲低组、多柔比星组、LINC01410敲低+多柔比星组,各组给予相应的慢病毒转染和多柔比星处理。采用实时荧光定量PCR法检测B-NHL细胞系中LINC01410 RNA的相对表达水平,采用细胞计数检测法检测各组B-NHL细胞的细胞活力,采用流式细胞术检测B-NHL细胞的凋亡情况,采用蛋白质免疫印迹法检测B-NHL细胞中多聚腺苷二磷酸-核糖聚合酶(PARP)蛋白的相对表达水平。结果共筛选出30个高风险LncRNA,风险比的范围为2.41~20.80,其中LINC01410的风险比为2.51。在Raji、Ramos细胞系中,慢病毒对照组、LINC01410敲低组、多柔比星组、LINC01410敲低+多柔比星组的细胞活力依次降低,细胞凋亡率依次升高(均P<0.05);多柔比星组与LINC01410敲低+多柔比星组的PARP蛋白相对表达水平均较慢病毒对照组升高,LINC01410敲低+多柔比星组的PARP蛋白相对表达水平较多柔比星组升高(均P<0.05)。LINC01410过表达的B-NHL患者的生存时间短于LINC01410低表达者(P<0.05)。结论LINC01410过表达提示B-NHL患者预后不良。LINC01410敲低可上调凋亡蛋白PARP的表达,促进B-NHL细胞凋亡,抑制B-NHL细胞活力,与多柔比星联用可产生增效协同作用。

Objective To explore the effects of long noncoding RNA(LncRNA)LINC01410 on the proliferation and apoptosis of B-cell non-Hodgkin′s lymphoma(B-NHL)cells.Methods The Cox regression model was used to screen the LncRNAs related to the prognosis of B-NHL in GEO dataset(GSE10846).B-NHL cell lines,Raji and Ramos,were cultured in vitro and randomly divided into lentivirus control group,LINC01410 knockdown group,doxorubicin group and LINC01410 knockdown+doxorubicin group.Each group was given corresponding lentivirus transfection and doxorubicin treatment.The relative expression level of LINC01410 RNA in B-NHL cell line was detected by real-time fluorescence quantitative PCR.The viability of B-NHL cells was detected in each group by cell counting kit-8 assay.The apoptosis of B-NHL cells was determined by flow cytometry.The relative expression level of poly(ADP-ribose)polymerase(PARP)protein in B-NHL cells was detected by Western blotting assay.Results A total of 30 high-risk LncRNAs were identified,and the hazard ratio ranged from 2.41 to 20.80,wherein the hazard ratio for LINC01410 was 2.51.In the Raji or Ramos cell line,cell viability decreased while the apoptosis rate increased in the lentivirus control group,LINC01410 knockdown group,doxorubicin group and LINC01410 knockdown+doxorubicin group successively(all P<0.05);the doxorubicin and LINC01410 knockdown+doxorubicin groups exhibited elevated relative expression levels of PARP protein than the lentivirus control group,and the LINC01410 knockdown+doxorubicin group yielded a higher relative expression level of PARP protein than the doxorubicin group(all P<0.05).B-NHL patients with LINC01410 overexpression had shorter survival time than those with low expression of LINC01410(P<0.05).Conclusion LINC01410 overexpression indicates a poor prognosis in B-NHL patients.LINC01410 knockdown might up-regulate the expression of apoptin PARP,promote the apoptosis of B-NHL cells,inhibit the viability of B-NHL cells,and might produce synergetic effects in combination with doxorubicin.