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. 2008 Jun 21;14(23):3754-8.
doi: 10.3748/wjg.14.3754.

Effect of fragile histidine triad gene transduction on proliferation and apoptosis of human hepatocellular carcinoma cells

Rong-Hua Xu  1 , Liang-Yan Zheng, Dong-Lei He, Jian Tong, Li-Ping Zheng, Wu-Ping Zheng, Jin Meng, Li-Ping Xia, Cong-Jun Wang, Ji-Lin Yi
Affiliations

Effect of fragile histidine triad gene transduction on proliferation and apoptosis of human hepatocellular carcinoma cells

Rong-Hua Xu et al. World J Gastroenterol. .

Abstract

Aim: To evaluate the inhibitory effects of human fragile histidine triad (FHIT) gene on cell proliferation and apoptosis in human hepatocellular carcinoma line Hep3B in vitro.

Methods: A recombinant pcDNA3.1 (+)/FHIT including the functional region of FHIT gene was constructed and transferred into human hepatocellular carcinoma cells in vitro. mRNA and protein expression of the FHIT gene in the transfected cells was detected by RT-PCR and Western blot, respectively. The effect of FHIT on proliferation was detected by MTT assay. Changes in cell cycle and apoptosis were assayed by flow cytometry. Five mice received subcutaneous transplantation of Hep3B-FHIT; 5 mice received subcutaneous transplantation of normal Hep3B and Hep3B-C as controls. The body weight of nude mice and tumor growth were measured.

Results: RT-PCR and Western blot analysis showed that the expression level of FHIT-mRNA and FHIT protein was higher in Hep3B cells after infection with pcDNA3.1 (+)/FHIT. The growth of Hep3B cells treated with pcDNA3.1 (+)/FHIT was significantly inhibited. The pcDNA3.1 (+)/FHIT-transfected Hep3B cells showed a significantly higher cell rate at G(0)-G(1) phase and increased apoptosis in comparison with controls (P < 0.05). The growth of transplanted tumor was inhibited markedly by FHIT. Tumors arising from the Hep3B-FHIT cells occurred much later than those arising from the Hep3B and Hep3B-C cells. The growth of Hep3B-FHIT cells was slow and the tumor volume was low.

Conclusion: Transduction of FHIT gene inhibits the growth of human hepatocellular carcinoma cells and induces cell apoptosis in vivo and in vitro.

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Figures

Figure 1
Figure 1
Identification of pcDNA3.1 (+)/FHIT by enzyme-cutting assay. M1: Marker λ/DNA/EcoRI+HindIII; M2: Marker DL2000, lane 1: pcDNA3.1 (+)/FHIT, lane 2: pcDNA3.1 (+)/FHIT post enzyme cutting.
Figure 2
Figure 2
Expression of FHIT mRNA and protein in Hep3B-FHIT, Hep3B-C and Hep3B cells. A: FHIT mRNA; B: FHIT protein. M: 1000 bp marker; Lane 1: Hep3B cells; lane 2: Hep3B-C cells; lane 3: Hep3B-FHIT cells.
Figure 3
Figure 3
Growth curves. A: Growth curves of human hepatocellular carcinoma cells. Luminous absorbance of Hep3B-FHIT, Hep3B-C and Hep3B cells was measured by ELISA (wavelength of 570 nm) after DMSO was added. The measurement was done once a day for 6 d; B: Growth curves of tumors after implantation of Hep3B, Hep3B-C or Hep3B-FHIT cells in nude mice. The mice were injected sc with 1 × 107 (0.15 mL/mouse) Hep3B, Hep3B-C or Hep3B-FHIT cells. After implantation, tumor growth was detected weekly.
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References

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