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Effects of HSP70 Antisense Oligonucleotide on the Proliferation and Apoptosis of Human Hepatocel_论文

J Huazhong Univ S ci Technol [Med Sci] 30(3):337-343,2010 DOI 10.1007/s11596-010-0353-4 J Huazhong Uni v Sci Technol [Med Sci] 30 (3): 2010 337 Effects of HSP70 Antisense Oligonucleotide on the Pr olifer ation and Apoptosis of Human Hepatocellular Car cinoma Cells Xue YANG (杨 雪)1, Haibin HE (贺海斌)2, Wei YANG (杨 威)1, Tao SONG (宋 涛)1, Cheng GUO (郭 成)1, Xin ZHENG (郑 鑫)1, Qingguang LIU (刘青光)1# 1 Depar tment of Hepa tobiliar y Sur ger y, the First Affiliated Hospita l, Medica l School of Xi’ Jiaotong Unive rsity, Xi’n 710061, an a China 2 Depar tment of Gener al Sur ger y, Ningbo Medical Trea tment Center Lihuili Hospita l, Ningbo 315040, China Huazhong University of Science and Technology and Springer-Verlag Berlin Heidelberg 2010 Summ ar y: The study investigated the effects of heat shock protein 70 (HSP70) antisense oligonucleotide (ASODN) on the proliferation and apoptosis of a human hepatocellular carcinoma cell line (SMMC-7721 cells) in vitro. HSP70 oligonucleotide was transfected into SMMC-7721 cells by the mediation of SofastTM transfection reagent. Inhibition rate of SMMC-7721 cells was determined by using MTT method. Apoptosis rate and cell cycle distribution were measured by flow cytometry. Immunocytochemistry staining was used to observe the expression of HSP70, Bcl-2 and Bax. The results showed that HSP70 ASODN at various concentrations could significantly inhibit the growth of SMMC-7721 cells, and the inhibition effect peaked 48 h after transfection with 400-nmol/L HSP70 ASODN. Cytometric analysis showed the apoptotic rate was increased in a dose- and time-dependent manner in the HSP70 ASODN-treated cells. The percentage of cells in the G 2/M and S phases was significantly decreased and that in the G0/G1 phase increa sed as the HSP70 ASODN concentration was elevated and the exposure time prolonged. Immunocytochemistry showed that treatment of SMMC-7721 cells with HSP70 ASODN resulted in decreased expressions of HSP70 and Bcl-2 proteins, and an increased expression of Bax protein. It was concluded that the HSP70 ASODN can inhibit the growth of the SMMC-7721 cells and increase cell apoptosis by down-regulating the expression of HSP70. HSP70 ASODN holds promise for the treatment of hepatocellular c arcinoma. Key wor ds: hepatocellular carcinoma; heat shock protein 70; antisense oligonucleotide; apoptosis Hepatocellular carcinoma (HCC) is one of the most common malignant diseases and has the lowest 5-year survival rate of any cancer. No satisfactor

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