[Inducing apoptosis of HeLa cell lines by transfection of c-myc antisense oligonucleotides].
Rongrong Li, Daoxin Ma, Yufeng Cheng
Abstract
Rongrong Li, Daoxin Ma, Yufeng Cheng
Abstract
OBJECTIVE: To investigate the effect on apoptosis of HeLa cells by transfecting c-myc antisense oligonucleotides and seek a new method to enhance radiosensitivity of tumor cells. METHODS: Immunohistochemistry staining and RT-PCR were conducted to identify the effect of ASODN. Giemsa staining, flow cytometry (FCM) and DNA ladder were performed to detect the apoptosis of HeLa cells. RESULTS: After transfection of ASODN (c-myc), the levels of both c-myc mRNA and C-MYC protein expression were reduced (P < 0.05). Giemsa staining demonstrated apoptosis-bodies on the c-myc ASODN-transfected cells. DNA ladder presented the typical ladder of apoptosis. The apoptosis rate of HeLa cells transfected with c-myc ASODN was (10.29 +/- 0.66)%, transfected c-myc ASODN combined irradiation was (16.83 +/- 0.57)%, which were much higher than that of control group (1.79 +/- 0.19)% (P < 0.05). CONCLUSION: The c-myc ASODN could effectively down-regulate the expression of c-myc; furthermore, transfection of c-myc ASODN could more effectively induce apoptosis of HeLa cells which subsequently enhanced radiosensitivity of HeLa cells.
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OBJECTIVE: To investigate the effect on apoptosis of HeLa cells by transfecting c-myc antisense oligonucleotides and seek a new method to enhance radiosensitivity of tumor cells. METHODS: Immunohistochemistry staining and RT-PCR were conducted to identify the effect of ASODN. Giemsa staining, flow cytometry (FCM) and DNA ladder were performed to detect the apoptosis of HeLa cells. RESULTS: After transfection of ASODN (c-myc), the levels of both c-myc mRNA and C-MYC protein expression were reduced (P < 0.05). Giemsa staining demonstrated apoptosis-bodies on the c-myc ASODN-transfected cells. DNA ladder presented the typical ladder of apoptosis. The apoptosis rate of HeLa cells transfected with c-myc ASODN was (10.29 +/- 0.66)%, transfected c-myc ASODN combined irradiation was (16.83 +/- 0.57)%, which were much higher than that of control group (1.79 +/- 0.19)% (P < 0.05). CONCLUSION: The c-myc ASODN could effectively down-regulate the expression of c-myc; furthermore, transfection of c-myc ASODN could more effectively induce apoptosis of HeLa cells which subsequently enhanced radiosensitivity of HeLa cells.
Key concepts: HeLa, Transfection, Apoptosis, Molecular biology, Oligonucleotide, Radiosensitivity, Flow cytometry, Biology