Effects of blocking Notch3 pathway by siRNA under hypoxic condition on proliferation of human lung adenocarcinoma A549 cells
Online published: 2013-05-03
Supported by
Shanghai Jinshan District Science and Technology Innovation Foundation, 2011-3-06; Shanghai Education Committee Foundation,11YZ44
Objective To observe the effects of blocking Notch3 by siRNA under hypoxic condition on the proliferation of human lung adenocarcinoma A549 cells. Methods Human lung adenocarcinoma A549 cells were cultured under hypoxic condition (1% O2). After transfection with Notch3 siRNA (Notch3 siRNA group), RT-PCR and Western blotting were used to detect the expression of Notch3 and its downstream gene HES1 mRNA and protein in A549 cells respectively, and the cell viability 24 h, 48 h and 72 h after transfection was determined by MTT assay. Besides, negative control group and blank control group were established. Results After transfection with Notch3 siRNA, the expression of Notch3 and HES1 mRNA and protein in A549 cells was significantly decreased (P<0.05). Transfection with Notch3 siRNA time-dependently inhibited the growth of A549 cells, and the cell viability 48 h and 72 h after transfection in Notch3 siRNA group was significantly different from those in negative control group and blank control group (P<0.05). Conclusion Transfection with Notch3 siRNA under hypoxic condition may effectively inhibit the expression of Notch3 and its downstream gene HES1 in human lung adenocarcinoma A549 cells, and may suppress the growth of A549 cells.
Key words: Notch3; lung adenocarcinoma; siRNA; hypoxia
HAN Rui-chao, FAN Zhi-wen, ZHOU Min . Effects of blocking Notch3 pathway by siRNA under hypoxic condition on proliferation of human lung adenocarcinoma A549 cells[J]. Journal of Shanghai Jiao Tong University (Medical Science), 2013 , 33(4) : 421 . DOI: 10.3969/j.issn.1674-8115.2013.04.008
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