Inhibitory effects of thioredoxin reductase antisense RNA on the growth of human hepatocellular carcinoma cells.
Thioredoxin reductase (TrxR) in conjunction with thioredoxin (Trx) is a ubiquitous intracellular oxidoreductase system with antioxidant and redox regulatory roles. In some human tumors, the thioredoxin system is found overexpressed. We have used an antisense approach to investigate whether inhibition of TrxR overexpression can suppress the growth of human hepatocellular carcinoma SMMC-7721 cells. TrxR cDNA fragment was inserted in the antisense direction into pcDNA3.1/myc-His and SMMC-7721 cells were stably transfected with the plasmid construct. The results showed that TrxR antisense RNA could significantly reduce TrxR mRNA level and activity, and suppress the growth of SMMC-7721 cells. Cell-cycle analysis showed G2/M phase arrest in SMMC-7721 cells transfected with TrxR antisense plasmid. TrxR antisense RNA could significantly increase p53 mRNA level and decrease Bcl-2 mRNA level by reverse transcriptase-polymerase chain reaction (RT-PCR). Furthermore a significant decrease in human telomerase reverse transcriptase (hTERT) mRNA level was found in SMMC-7721 cells transfected with TrxR antisense plasmid. Flow cytometry and telomere fluorescence in situ hybridization (Flow FISH) showed that TrxR antisense RNA could significantly reduce the telomere fluorescence in SMMC-7721 cells. The results suggested that TrxR antisene RNA inhibited the growth of SMMC-7721 cells through an accumulation of cell cycle at G2/M phase, an increase in p53 mRNA level and a reduction in telomere fluorescence and Bcl-2, hTERT mRNA levels.
Gan
Lu
L
College of Life Science and Technology, Huazhong University of Science and Technology, Wuhan 430074,China.
Yang
Xiang-Liang
XL
Liu
Qiong
Q
Xu
Hui-Bi
HB
D006528:Carcinoma, Hepatocellular / Q000201:enzymology* / Q000473:pathology; D002453:Cell Cycle / Q000502:physiology; D045744:Cell Line, Tumor; D049109:Cell Proliferation; D002470:Cell Survival; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism; D006801:Humans; D008113:Liver Neoplasms / Q000201:enzymology* / Q000473:pathology; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000235:genetics / Q000378:metabolism; D016372:RNA, Antisense / Q000235:genetics / Q000378:metabolism*; D019098:Telomerase / Q000235:genetics / Q000378:metabolism; D016615:Telomere / Q000378:metabolism; D013880:Thioredoxin-Disulfide Reductase / Q000235:genetics* / Q000378:metabolism; D016159:Tumor Suppressor Protein p53 / Q000235:genetics / Q000378:metabolism
D004268:DNA-Binding Proteins; D019253:Proto-Oncogene Proteins c-bcl-2; D016372:RNA, Antisense; D016159:Tumor Suppressor Protein p53; D013880:Thioredoxin-Disulfide Reductase; D019098:Telomerase