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[Experimental therapy of survivin antisense oligonucleotide for human ovarian cancer cell SKOV3].

Shu Qin Wei, Sheng Bi, Jian-Hua Zheng, Guangmei Zhang, Sui Li-hua, Shangha Pan

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Abstract

BACKGROUND & OBJECTIVE: Survivin abnormally overexpresses in a variety of human tumors, it may play an important role in the development of tumor. This study was designed to investigate the inhibitory effects of survivin antisense oligonucleotide (ASODN) on human ovarian carcinoma cell SKOV3 in vivo and its mechanism. METHODS: SKOV3 cells were transfected with survivin ASODN mediated by DOTAP liposome reagent, MTT method was used to observe the inhibitory rate, Western blot analysis was used to determine relating gene expression. Flow cytometry, DNA ladder analysis, and DAPI staining were used to examine cell apoptosis. Kinase activity test was used to examine the changes of caspase-3 activity. RESULTS: The expression of survivin in SKOV3 cells decreased after transfected with survivin ASODN. Survivin ASODN has obviously inhibited the growth of SKOV3 cells after transfection. The inhibitory rate of 1000 ng/ml ASODN transfection group was (60.30+/-2.95)%, which is obviously higher than control group (P< 0.05). Survivin ASODN transfection induced cell cycle arrest in G1 phrase, while the apoptotic rate increased from 0.65% to 32.10%. Caspase-3 activity (0.998+/-0.001) increased significantly after transfected with survivin ASODN (P< 0.01). CONCLUSION: Survivin ASODN,which can inhibit human ovarian carcinoma SKOV3 cells proliferation and induce apoptosis, is a prospective anti-cancer drug.

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What this paper is about

BACKGROUND & OBJECTIVE: Survivin abnormally overexpresses in a variety of human tumors, it may play an important role in the development of tumor. This study was designed to investigate the inhibitory effects of survivin antisense oligonucleotide (ASODN) on human ovarian carcinoma cell SKOV3 in vivo and its mechanism. METHODS: SKOV3 cells were transfected with survivin ASODN mediated by DOTAP liposome reagent, MTT method was used to observe the inhibitory rate, Western blot analysis was used to determine relating gene expression. Flow cytometry, DNA ladder analysis, and DAPI staining were used to examine cell apoptosis. Kinase activity test was used to examine the changes of caspase-3 activity. RESULTS: The expression of survivin in SKOV3 cells decreased after transfected with survivin ASODN. Survivin ASODN has obviously inhibited the growth of SKOV3 cells after transfection. The inhibitory rate of 1000 ng/ml ASODN transfection group was (60.30+/-2.95)%, which is obviously higher than control group (P< 0.05). Survivin ASODN transfection induced cell cycle arrest in G1 phrase, while the apoptotic rate increased from 0.65% to 32.10%. Caspase-3 activity (0.998+/-0.001) increased significantly after transfected with survivin ASODN (P< 0.01). CONCLUSION: Survivin ASODN,which can inhibit human ovarian carcinoma SKOV3 cells proliferation and induce apoptosis, is a prospective anti-cancer drug.

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Available abstract

BACKGROUND & OBJECTIVE: Survivin abnormally overexpresses in a variety of human tumors, it may play an important role in the development of tumor. This study was designed to investigate the inhibitory effects of survivin antisense oligonucleotide (ASODN) on human ovarian carcinoma cell SKOV3 in vivo and its mechanism. METHODS: SKOV3 cells were transfected with survivin ASODN mediated by DOTAP liposome reagent, MTT method was used to observe the inhibitory rate, Western blot analysis was used to determine relating gene expression. Flow cytometry, DNA ladder analysis, and DAPI staining were used to examine cell apoptosis. Kinase activity test was used to examine the changes of caspase-3 activity. RESULTS: The expression of survivin in SKOV3 cells decreased after transfected with survivin ASODN. Survivin ASODN has obviously inhibited the growth of SKOV3 cells after transfection. The inhibitory rate of 1000 ng/ml ASODN transfection group was (60.30+/-2.95)%, which is obviously higher than control group (P< 0.05). Survivin ASODN transfection induced cell cycle arrest in G1 phrase, while the apoptotic rate increased from 0.65% to 32.10%. Caspase-3 activity (0.998+/-0.001) increased significantly after transfected with survivin ASODN (P< 0.01). CONCLUSION: Survivin ASODN,which can inhibit human ovarian carcinoma SKOV3 cells proliferation and induce apoptosis, is a prospective anti-cancer drug.

Key concepts: Survivin, Transfection, Apoptosis, Molecular biology, DAPI, Cell cycle, Cancer research, Cell growth

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