Objective.. To study induction of apoptosis and inhibition of proliferation in Hep-2 by antisense survivin RNA. Methods： Antisense survivin RNA expression vector was constructed and then was transfected to human lary...Objective.. To study induction of apoptosis and inhibition of proliferation in Hep-2 by antisense survivin RNA. Methods： Antisense survivin RNA expression vector was constructed and then was transfected to human laryngeal carcinoma cell line Hep-2 by lipofectamine. HpEGFP/survivin cells （transfected with the combinant of antisense survivin RNA） were obstained by using G418. The levels of survivin protein before and after transfection were determined by Western-blot. Proliferation activity was measured by MTT assay. The experiment of colony formation in soft agar was carried out for assessing ability of proliferation of Hep-2 cell. Apoptosis was assessed by flow cytometry and acrdine orange（AO）. Results： After antisense survivin RNA plasmids were transfected, the level of survivin protein was inhibited in Hep-2. Compared with control, proliferation of HpEGFP/survivin cells were suppressed significantly. The experiment of colony formation in soft agar showed the ability of colony formation decreased in HpEGFP/survivin cells compared to control （P〈0.05）. Apoptosis rate increased about 1.81-folds compared with control. Conclusion.. The antisense survivin RNA can partly inhibit the level of surviivin protein expression in Hep-2 and can induce apoptosis and inhibit the proliferation of Hep-2 by down-regulating the expression of endogenous survivin in vitro.展开更多
基金Surpported by Bureau of Science and Technology of Changchun City (03-180S19)
文摘Objective.. To study induction of apoptosis and inhibition of proliferation in Hep-2 by antisense survivin RNA. Methods： Antisense survivin RNA expression vector was constructed and then was transfected to human laryngeal carcinoma cell line Hep-2 by lipofectamine. HpEGFP/survivin cells （transfected with the combinant of antisense survivin RNA） were obstained by using G418. The levels of survivin protein before and after transfection were determined by Western-blot. Proliferation activity was measured by MTT assay. The experiment of colony formation in soft agar was carried out for assessing ability of proliferation of Hep-2 cell. Apoptosis was assessed by flow cytometry and acrdine orange（AO）. Results： After antisense survivin RNA plasmids were transfected, the level of survivin protein was inhibited in Hep-2. Compared with control, proliferation of HpEGFP/survivin cells were suppressed significantly. The experiment of colony formation in soft agar showed the ability of colony formation decreased in HpEGFP/survivin cells compared to control （P〈0.05）. Apoptosis rate increased about 1.81-folds compared with control. Conclusion.. The antisense survivin RNA can partly inhibit the level of surviivin protein expression in Hep-2 and can induce apoptosis and inhibit the proliferation of Hep-2 by down-regulating the expression of endogenous survivin in vitro.
