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siRNA 下调 β-1,3-N-乙酰氨基葡萄糖基转移酶-8 抑制人胃癌的生长。

Down-regulation of β-1,3-N-acetylglucosaminyltransferase-8 by siRNA inhibits the growth of human gastric cancer.

机构信息

Department of Biochemistry and Molecular Biology, Soochow University, Suzhou, Jiangsu 215123, PR China.

出版信息

Mol Med Rep. 2011 May-Jun;4(3):497-503. doi: 10.3892/mmr.2011.448. Epub 2011 Mar 4.

Abstract

β-1,3-N-acetylglucosaminyltransferase-8 (β3Gn-T8) is the most recently identified enzyme in the β3Gn-T family, but its biological function is poorly understood. To elucidate the effects of β3Gn-T8 on gastric cancer behavior, β3Gn-T8 was down-regulated in AGS cells using small interfering RNA (siRNA). The mRNA and protein expression levels of β3Gn-T8 were detected using RT-PCR and Western blotting, respectively, and sequence-specific inhibition using siRNA was also measured using RT-PCR in human SPCA-1 and SGC-7901 cells. The cell proliferation rate was determined using MTT and the percentage of apoptotic cells was measured using flow cytometry. AGS cells transfected with β3Gn-T8 siRNA were subcutaneously transplanted into nude mice and tumorigenicity was assessed. The siRNA efficiently suppressed β3Gn-T8 expression in AGS cells, and the down-regulation of β3Gn-T8 caused significant inhibition of tumor cell growth in vitro. The apoptotic rate of AGS cells increased to 10.13% 48 h after siRNA transfection, which was five times that of the control cells. Furthermore, the knockdown of β3Gn-T8 expression reduced the tumorigenicity of gastric cancer cells in nude mice, suggesting that β3Gn-T8 has potential as a gastric cancer therapeutic target.

摘要

β-1,3-N-乙酰氨基葡萄糖基转移酶-8(β3Gn-T8)是β3Gn-T 家族中最近发现的酶,但它的生物学功能知之甚少。为了阐明β3Gn-T8 对胃癌行为的影响,使用小干扰 RNA(siRNA)下调 AGS 细胞中的β3Gn-T8。使用 RT-PCR 和 Western blot 分别检测β3Gn-T8 的 mRNA 和蛋白表达水平,并用 RT-PCR 测量人 SPCA-1 和 SGC-7901 细胞中 siRNA 的序列特异性抑制。使用 MTT 测定细胞增殖率,并用流式细胞术测定凋亡细胞的百分比。将转染了β3Gn-T8 siRNA 的 AGS 细胞皮下移植到裸鼠中,并评估其致瘤性。该 siRNA 有效地抑制了 AGS 细胞中β3Gn-T8 的表达,β3Gn-T8 的下调导致体外肿瘤细胞生长的显著抑制。siRNA 转染 48 小时后,AGS 细胞的凋亡率增加到 10.13%,是对照细胞的五倍。此外,β3Gn-T8 表达的敲低降低了裸鼠中胃癌细胞的致瘤性,表明β3Gn-T8 可能成为胃癌的治疗靶点。

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