Expression of Twist gene in human hepatocellular carcinoma cell strains of different metastatic potential

Abstract

In order to investigate the role of Twist gene in the metastasis of hepatocellular carcinoma (HCC), total RNA was respectively extracted from three HCC cell strains with different metastatic potentials, HepG2, MHCC-97L and MHCC-97H. The first strand cDNA was synthesized by reverse transcription, which was then used as template to perform fluorescent quantitative polymerase chain reaction (FQ-PCR). The quantity of Twist gene expression was normalized by that of the housekeeping gene, GAPDH for each sample. ANOVA was used to estimate the relationship between Twist gene and metastasis potential of HCC. The results showed that the normalized initial cDNA concentrations of Twist gene in HepG2, MHCC-97L and MHCC-97H were (9.45±0.25)×10⁻⁴, (1.82±0.41)×10⁻³, (3.06±0.62)×10⁻³, respectively. FQ-PCR revealed significant differences in the expression level of Twist among HCC cell strains with different metastatic potentials. It was concluded that high expression level of Twist was closely associated with more aggressive behaviors of HCC. Twist provides a novel indicator for HCC metastasis.

Keywords

hepatocellular carcinoma / Twist gene / metastasis / fluorescent polymerase chain reaction

References

[1] ThieryJ. P.. Epithelial-mesenchymal transitions in tumor progression. Nat Rev Cancer, 2002, 2: 442-454

[2] KuperH., YeW., BroomeU., et al.. The risk of liver and bile duct cancer in patients with chronic viral hepatitis, alcoholism, or cirrhosis. Hepatology, 2001, 34: 714-718

[3] ChambersA. F., GroomA. C., MacDonaldI. C.. Dissemination and growth of cancer cells in metastatic sites. Nat Rev Cancer, 2002, 2: 563-572

[4] ThieryJ. P.. Epithelial-mesenchymal transitions in development and pathologies. Curr Opin Cell Biol, 2003, 15: 740-746

[5] YibinK., JoanM.. Epithelial-mesenchymal transitions: twist in development and metastasis. Cell, 2004, 118: 277-279

[6] JingY., SenduralA. M., JoanaL. D., et al.. Twist, a master regulator of morphogenesis, plays an essential role in tumor metastasis. Cell, 2004, 117: 927-939

[7] SugimuraM., DonatoR., KakkarV. V., et al.. Annexin V as a probe of the contribution of anionic phospholipids to the procoagulant activity of tumor cell surfaces. Blood Coagul Fibrin, 1994, 5(3): 365-373

[8] LiY., TangZ. Y., YeS. L., et al.. Establishment of cell clones with different metastatic potential from the metastatic hepatocellular carcinoma cell line MHCC97. World J Gastroenterol, 2001, 7(5): 630-636

[9] TianJ., TangZ. Y., YeS. L., et al.. New human hepatocellular carcinoma (HCC) cell line with highly metastatic potential (MHCC97) and its expressions of the factors associated with metastasis. Br J Cancer, 1999, 81(5): 814-821

[10] LiY., TangZ. Y., YeS. L., et al.. Isolation and establishment of human hepatocellular carcinoma cell clones with various metastatic potentials. Chin J Hepatobiliary Surg, 2002, 7(11): 681-685

[11] MehrotraJ., ValiM., McVeighM., et al.. Very high frequency of hypermethylated genes in breast cancer metastasis to the bone, brain, and lung. Clin Cancer Res, 2004, 10: 3104-3109