Zheng Ruiji, Huang Yiqun, Ma Xudong
Department of Hematology, Zhangzhou Affiliated Hospital of Fujian Medical University, Zhangzhou 363000, Fujian Province, China.
Zhonghua Xue Ye Xue Za Zhi. 2015 Jan;36(1):49-52. doi: 10.3760/cma.j.issn.0253-2727.2015.01.012.
To investigate the effect of silencing mTOR gene by RNA interference on proliferation and apoptosis, and its mechanism on mantle cell lymphoma Jeko-1 cell Line.
The hairpin-like oligonucleotide sequences targeting mTOR gene was designed and transfected into Jeko-1 cells by lipofectamine TM 2000. The mTOR mRNA and protein were detected by RQ-PCR and Western blot. Cell growth was determined by MTT. Cell apoptosis was analyzed by flow cytometry. The expressions of Bcl-2, Bax, procaspase-3, procaspase-9, P70S6K,and p-P70S6K were detected by Western blot.
mTOR mRNA was markedly suppressed by shRNA targeting mTOR. mTOR shRNA suppressed proliferation and induced cells apoptosis of Jeko-1 cells. The cell apoptotic rates were (36.62 ± 3.24)%, (2.58 ± 1.04)%, (1.24 ± 0.30)% respectively, in mTOR shRNA, Neg-shRNA and Blank with statistically significant difference among them (P<0.05). mTOR shRNA down-regulated the expressions of Bcl-2, proCaspase3, proCaspase9 and p-70S6K, up-regulated the expression of Bax.
Deplete of mTOR gene may be realized through inhibiting the Akt/mTOR signaling pathway to promote the cell apoptosis and inhibit cell growth in Jeko-1 cell line.
探讨RNA干扰沉默mTOR基因对套细胞淋巴瘤Jeko-1细胞株增殖和凋亡的影响及其机制。
设计针对mTOR基因的发夹状寡核苷酸序列，通过脂质体TM 2000转染至Jeko-1细胞。采用RQ-PCR和Western blot检测mTOR mRNA和蛋白。用MTT法测定细胞生长情况。通过流式细胞术分析细胞凋亡。用Western blot检测Bcl-2、Bax、procaspase-3、procaspase-9、P70S6K和p-P70S6K的表达。
靶向mTOR的shRNA显著抑制mTOR mRNA。mTOR shRNA抑制Jeko-1细胞的增殖并诱导其凋亡。mTOR shRNA、Neg-shRNA和空白组的细胞凋亡率分别为(36.62±3.24)%、(2.58±1.04)%、(1.24±0.30)%，差异有统计学意义(P<0.05)。mTOR shRNA下调Bcl-2、proCaspase3、proCaspase9和p-70S6K的表达，上调Bax的表达。
在Jeko-1细胞株中，通过抑制Akt/mTOR信号通路可能实现mTOR基因缺失，从而促进细胞凋亡并抑制细胞生长。
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