LncRNA MALAT1 promotes tumorigenesis and immune escape of diffuse large B cell lymphoma by sponging miR-195
LncRNA MALAT1 promotes tumorigenesis and immune escape of diffuse large B cell lymphoma by sponging miR-195
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DOI：
10.1016/j.lfs.2019.03.040
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发表时间：
2019-08-15
期刊：
影响因子：
6.1
通讯作者：
Gong, Yi
中科院分区：
文献类型：
--
作者：
Wang, Qing-Ming;Lian, Guang-Yu;Gong, Yi
背景：PD-L1增强肿瘤的发生和免疫逃逸能力。PD-L1调控弥漫性大B细胞淋巴瘤(DLBCL)发生和免疫逃逸的上游机制尚不清楚。方法：采用人DLBCL细胞株OCI-Ly10和DLBCL患者标本。MALAT1被shRNA击倒。MiR-195抑制剂对MIR-195有抑制作用。RT-qPCR检测MALAT1、PD-L1、miR-195和CD8水平。免疫印迹法检测PD-L1、RAS、p-ERK1/2、ERK1/2、slug、E-cadherin、N-cadherin、Vimentin的蛋白水平。用荧光素酶检测MALAT1与miR-195、miR-195与PDL1的相互作用。MIT法检测OCI-Ly10细胞增殖；Annexin V/PI法检测细胞凋亡率。Transwell法检测细胞的迁移能力。用乳酸脱氢酶细胞毒试剂盒检测CD8(+)T细胞的细胞毒作用。结果：DLBCL组织中MALAT1、PD-L1和CD8表达上调，miR-195表达下调。MIR-195与MALAT1、PD-L1呈负相关。MALAT1可通过海绵结合miR-195来调节PD-11的表达。ShMALAT1治疗可提高miR-195水平，降低PD-LL水平。抑制细胞增殖、迁移和免疫逃逸能力，增加OCI-Ly10细胞凋亡率。ShMALAT1对OCI-Ly10细胞也有促进CD8(+)T细胞增殖和抑制细胞凋亡的作用。MALAT1基因的敲除也通过RAS/ERK信号通路抑制了EMT样信号转导。结论：长非编码RNA MALAT1融合miR-195，通过调节PD-L1调节DLBCL的增殖、凋亡、迁移和免疫逃逸能力。
Background: PD-L1 enhanced the tumorigenesis and immune escape abilities of cancers. The upstream mechanisms of PD-L1 in regulating tumorigenesis and immune escape of diffuse large B cell lymphoma (DLBCL) remained unclear.Methods: Human DLBCL cell line OCI-Ly10 and DLBCL patient samples were used in this study. MALAT1 was knocked down by shRNA. MiR-195 was inhibited by miR-195 inhibitor. Levels of MALAT1, PD-L1, miR-195 and CD8 were detected by RT-qPCR. Protein levels of PD-L1, Ras, p-ERK1/2, ERK1/2, Slug, E-cadherin, N-cadherin, Vimentin were detected by western blotting. The interaction between MALAT1 and miR-195, miR-195 and PDL1 were detected by luciferase assay. OCI-Ly10 cell proliferation and apoptosis were detected by MIT and Annexin V/PI assays, respectively. Migration was detected by transwell assay. Cytotoxicity of CD8(+) T cells was detected by LDH cytotoxicity kit. Proliferation and apoptosis of CD8(+) T cell co-cultured with OCI-Ly10 cells were analyzed by CFSE and Annexin V/PI staining.Results: MALAT1, PD-L1 and CD8 were up-regulated in DLBCL tissues while miR-195 was down-regulated. MiR-195 was negatively correlated with MALAT1 and PD-L1. MALAT1 could sponge miR-195 to regulate the expression of PD-Ll. shMALAT1 treatment increased miR-195 level and decreased PD-Ll level. It also inhibited cell proliferation, migration and immune escape ability while increased apoptosis ratio of OCI-Ly10 cells. shMALAT1 treatment in OCI-Ly10 cells also promoted proliferation and inhibited apoptosis of CD8(+) T cells. Knocking down of MALAT1 also suppressed EMT-like process via Ras/ERK signaling pathway. These effects were all rescued by miR-195 inhibitor.Conclusion: Long non-coding RNA MALAT1 sponged miR-195 to regulate proliferation, apoptosis and migration and immune escape abilities of DLBCL by regulation of PD-L1.