The expression of octamer binding factor 4 （Oct4） gene in bladder cancer cell line T24 and its effects on the biological characteristics of the cells were investigated. RT-PCR and Western blot were employed to detec...The expression of octamer binding factor 4 （Oct4） gene in bladder cancer cell line T24 and its effects on the biological characteristics of the cells were investigated. RT-PCR and Western blot were employed to detect the expression of Oct4 in T24 cells. The changes of biological characteristics in T24 cells were analyzed before and after gene-silencing by Boyden chamber and MTT. The results showed that the expression of Oct4 gene was detectable in T24 cells by RT-PCR and Western blot. The expression of Oct4 gene and protein was down-regulated by siRNA, and average number of transwell cells in interference group, negative control group and blank control group was 101.40±54.56, 104.20± 10.03 and 111.00±11.90, respectively. There was significant difference in the proliferation abilit,） of the cells from 48 h, 72 h to 96 h after the interference by siRNA between interference group and negative group or blank control group （P〈0.05）. It was suggested that Oct4 gene was related with proliferation ability of T24 cells, but not with invasive capability.展开更多
Aim:Mitomycin C(MMC)is a commonly used as intravesical treatment for superficial bladder cancer.However,its role in combination with ras inhibition has not been investigated.The aim of this study was to explore the ro...Aim:Mitomycin C(MMC)is a commonly used as intravesical treatment for superficial bladder cancer.However,its role in combination with ras inhibition has not been investigated.The aim of this study was to explore the role of ras in MMC-induced apoptosis in T24 bladder cancer cells and to determine the efficacy of combination therapy in vitro.Methods:We measured the effects of various doses of MMC on apoptosis induction as well as on ras,ERK and Ki-67 protein expression by T24 cell line using immunocytochemistry,flow cytometry and Western blotting.We also tested the effect of siRNA on ras employed singly or in combination with MMC.Results:T24 cells expressed high level of ras protein.MMC treatment increased the level of ras and ERK protein expression after 24 h,and decreased these levels after 72 h.Ras siRNA(100 nmol/L)caused massive apoptosis associated with a marked decrease in ras expression in T24 cells.When combined with low doses of MMC,ras siRNA(50 nmol/L)sensitized T24 cells to apoptosis and decreased their expression of ras.The effect of combined therapy was higher than that of either compound used alone.Expression levels of ERK,a downstream target of ras,declined following combination therapy.Conclusion:Ras siRNA in combination with low dose MMC is a possible treatment strategy for patients with ras-positive bladder tumors.展开更多
文摘The expression of octamer binding factor 4 （Oct4） gene in bladder cancer cell line T24 and its effects on the biological characteristics of the cells were investigated. RT-PCR and Western blot were employed to detect the expression of Oct4 in T24 cells. The changes of biological characteristics in T24 cells were analyzed before and after gene-silencing by Boyden chamber and MTT. The results showed that the expression of Oct4 gene was detectable in T24 cells by RT-PCR and Western blot. The expression of Oct4 gene and protein was down-regulated by siRNA, and average number of transwell cells in interference group, negative control group and blank control group was 101.40±54.56, 104.20± 10.03 and 111.00±11.90, respectively. There was significant difference in the proliferation abilit,） of the cells from 48 h, 72 h to 96 h after the interference by siRNA between interference group and negative group or blank control group （P〈0.05）. It was suggested that Oct4 gene was related with proliferation ability of T24 cells, but not with invasive capability.
文摘Aim:Mitomycin C(MMC)is a commonly used as intravesical treatment for superficial bladder cancer.However,its role in combination with ras inhibition has not been investigated.The aim of this study was to explore the role of ras in MMC-induced apoptosis in T24 bladder cancer cells and to determine the efficacy of combination therapy in vitro.Methods:We measured the effects of various doses of MMC on apoptosis induction as well as on ras,ERK and Ki-67 protein expression by T24 cell line using immunocytochemistry,flow cytometry and Western blotting.We also tested the effect of siRNA on ras employed singly or in combination with MMC.Results:T24 cells expressed high level of ras protein.MMC treatment increased the level of ras and ERK protein expression after 24 h,and decreased these levels after 72 h.Ras siRNA(100 nmol/L)caused massive apoptosis associated with a marked decrease in ras expression in T24 cells.When combined with low doses of MMC,ras siRNA(50 nmol/L)sensitized T24 cells to apoptosis and decreased their expression of ras.The effect of combined therapy was higher than that of either compound used alone.Expression levels of ERK,a downstream target of ras,declined following combination therapy.Conclusion:Ras siRNA in combination with low dose MMC is a possible treatment strategy for patients with ras-positive bladder tumors.
