Guo Meng-Xian, Zou Yong, Lin Lu-Hui, Ma Xu-Dong, Huang Yi-Qun
Department of Hematology,Zhangzhou Municipal Hospital Affiliated to Fujian Medical University,Zhangzhou 363000,Fujian Province,China.
Department of Hematology,Zhangzhou Municipal Hospital Affiliated to Fujian Medical University,Zhangzhou 363000,Fujian Province,China. E-mail:
Zhongguo Shi Yan Xue Ye Xue Za Zhi. 2018 Jun;26(3):772-778. doi: 10.7534/j.issn.1009-2137.2018.03.023.
To investigate the effect of silencing NSD2 gene by RNA interference on the proliferation, apoptosis and the alteration of Akt /mTOR signaling pathway in diffuse large B cell lymphoma OCI-Ly3 cells.
The shRNA targeting NSD2 gene was transfected into OCI-Ly3 cells by lentivirus infection. The NSD2 mRNA and protein were detected by real time Q-PCR and Western blot, respectively. The cell proliferation was detected by CCK-8 and apoptosis was measured by flow cytometry. The expressions of BCL-2, BAX, caspase-3, Akt, p-Akt, p-mTOR, p-P70S6K, H3K36me2 were detected by Western blot.
After transfecting the OCI-Ly3 cells by NSD2-shRNA for 72 h, the expressions of NSD2 mRNA and protein both were down-regulated(P<0.05), the proliferation rate of cells in NSD2 shRNA group was significantly lower than that in control and Neg shRNA groups (P<0.05); the apoptosis rate of cells in NSD2 shRNA group was significantly higher than that in control and neg-shRNA group (30.37±4.22)% vs 1.36±0.52 % and 2.17±1.43)%(P<0.05); the expressions of BAX and caspase-3 were up-regulated, while the expression of BCL-2 was down-regulated; the H3K36me2 level significantly decreased as compared with control group, no obvious decrease of the total protein level of AKT was found, but the expressions of p-Akt, p-mTOR and p-70S6K were down-regulated.
The silencing NSD2 gene can inhibit the proliferation and induce the apoptosis of OCI-Ly3 cells, their mechanisms may relate with regulating the H3K36me2 level, specifically inhibiting the activivty of AKT/mTOR signal pathway.
探讨RNA干扰沉默NSD2基因对弥漫性大B细胞淋巴瘤OCI-Ly3细胞增殖、凋亡及Akt/mTOR信号通路变化的影响。
通过慢病毒感染将靶向NSD2基因的shRNA转染至OCI-Ly3细胞。分别采用实时荧光定量PCR和蛋白质免疫印迹法检测NSD2 mRNA和蛋白。采用CCK-8法检测细胞增殖，流式细胞术检测细胞凋亡。通过蛋白质免疫印迹法检测BCL-2、BAX、caspase-3、Akt、p-Akt、p-mTOR、p-P70S6K、H3K36me2的表达。
NSD2-shRNA转染OCI-Ly3细胞72小时后，NSD2 mRNA和蛋白表达均下调（P<0.05），NSD2 shRNA组细胞增殖率显著低于对照组和阴性对照shRNA组（P<0.05）；NSD2 shRNA组细胞凋亡率显著高于对照组和阴性对照shRNA组（30.37±4.22）% 比 1.36±0.52 % 和 2.17±1.43）%（P<0.05）；BAX和caspase-3表达上调，而BCL-2表达下调；与对照组相比，H3K36me2水平显著降低，未发现AKT总蛋白水平明显下降，但p-Akt、p-mTOR和p-70S6K表达下调。
沉默NSD2基因可抑制OCI-Ly3细胞增殖并诱导其凋亡，其机制可能与调节H3K36me2水平、特异性抑制AKT/mTOR信号通路活性有关。
