lncRNA XIST通过miR-375-Notch1轴调控急性T淋巴细胞白血病细胞增殖、侵袭的实验研究

目的 探讨lncRNA XIST通过miR-375-Notch1轴调控急性淋巴细胞白血病(T-acute lymphocytic leukemia,T-ALL)细胞增殖和侵袭的影响。方法 以2018年6月—2021年3月收治的57例T-ALL患者为观察对象(T-ALL组),另选55例同期健康体检者为对照组。qRT-PCR检测T-ALL组和对照组及T-ALL T淋巴细胞CCRF-CEM细胞株中lncRNA XIST水平;对CCRF-CEM细胞转染si-lncRNA XIST以沉默lncRNA XIST、转染miR-375模拟物以过表达miR-375;采用CCK-8法、Transwell侵袭实验法及流式细胞术检测CCRF-CEM细胞的增殖、侵袭及凋亡情况;荧光素酶报告基因方法检测lncRNA XIST靶基因;Western blot检测Notch1、ADAM-17、Hes1蛋白表达。结果 与对照组比较,T-ALL组的lncRNA XIST、Notch1 mRNA表达明显增加(P＜0.05),而miR-375水平明显下降(P＜0.05),且T-ALL组患者Notch1蛋白水平明显高于对照组(P＜0.001)。转染si-lncRNA XIST沉默lncRNA XIST后,CCRF-CEM细胞增殖数量、侵袭能力明显下降(P＜0.05),细胞凋亡明显增加(P＜0.05)。荧光素酶报告基因显示miR-375是lncRNA XIST的靶点。CCRF-CEM细胞转染miR-375后,细胞增殖数量、侵袭能力明显下降(P＜0.05),细胞凋亡明显增加(P＜0.05)。转染si-lncRNA XIST可以明显上调miR-375水平、下调Notch1 mRNA水平,同时下调Notch1、ADAM-17、Hes1蛋白表达。结论 lncRNA XIST在T-ALL患者中呈高表达,可能通过miR-375-Notch1轴影响白血病细胞生物学功能。

lncRNA XIST regulates the proliferation and invasion of acute T lymphocytic leukemia through the miR-375-Notch1 axis

Objective The aim of this study was to investigate the effect of lncRNA XIST on the proliferation and invasion of T-acute lymphocytic leukemia(T-ALL)cells through the regulation of miR-375-Notch1 axis. Methods Fifty-seven patients with T-ALL who were treated from June 2018 to March 2021 were collected as the observation object(T-ALL group),and 55 healthy people who underwent physical examination during the same period were selected as the control group.The levels of lncRNA XIST in the two groups of patients and CCRF-CEM cell lines were detected by qRT-PCR;CCRF-CEM cells were transfected with si-lncRNA XIST to silence lncRNA XIST,and miR-375 mimics were transfected to overexpress miR-375.CCK-8 method,transwell invasion test and flow cytometry were used to detect the proliferation,invasion and apoptosis of CCRF-CEM cells;the luciferase reporter gene method was used to detect lncRNA XIST target gene;Western blot was used to detect the expressions of Notch1,ADAM-17 and Hes1 protein. Results Compared with the control group,the expression of lncRNA XIST and Notch1 mRNA in the T-ALL group was significantly increased(P＜0.05),while the level of miR-375 was significantly decreased(P＜0.05),and the level of Notch1 protein in T-ALL patients was significantly higher than that in the control group(P＜0.001).After transfection with si-lncRNA XIST to silence lncRNA XIST,the proliferation and invasion of CCRF-CEM cells were significantly decreased(P＜0.05),and the apoptosis was significantly increased(P＜0.05).The luciferase reporter gene revealed that miR-375 was the target of lncRNA XIST.After CCRF-CEM cells were transfected with miR-375,the number of cell proliferation and invasion were significantly decreased(P＜0.05),and the apoptosis was significantly increased(P＜0.05).Transfection of si-lncRNA XIST could significantly up-regulate the level of miR-375,down-regulate the level of Notch1 mRNA,and down regulate the expression of Notch1,ADAM-17 and Hes1 protein(P＜0.05). Conclusion lncRNA XIST is highly expressed in T-ALL patients,which may affect the biological function of leukemia cells through the miR-375-Notch1 axis.