Abstract:

[Abstract]   Objective  To explore the effect of RNA interferencetargeting hypoxia-inducible factor-1α(HIF-1α)
gene on the proliferation of human tongue cancer cells (Tca8113and Tscca).  Methods  HIF-1α- specific short hairpin
RNA (shRNA) expression vector and empty shRNA vector were respectively transferred into two kinds of cultured human tongue cancer cells. The expression of HIF-1αgene was detected by RT-PCR and Western blot method. The clone formation assay was used to observe the cell proliferation. The cell counting was used to monitor the number of living cells. Flow cytometry was applied to detect cell cycle distribution. Additionally, the cells were injected into nude mice, and tumor forming condition was observed.   Results   After the shRNA HIF-1αplasmid was transfected into tongue cancer cells, the expressions of both HIF-1αmRNA and protein were significantly decreased in tongue cancer cells under hypoxia condition. The number of colony formation was significantly lower in HIF-1αsmall interfering RNA (siRNA) group than that in empty and negative control group. The cell counting revealed that the number of living cells was remarkably lower in HIF-1αsiRNA groupthan that in negative control group. Also, the cell cycle in HIF-1αsiRNA group was arrested at G0/G1phase after transfection and 48hours hypoxia culture. Besides, the tumor volume wasremarkably smaller in HIF-1αsiRNA groupthan that in negative control group after thetongue cancer cells transplanted into nude mice for several days. The statistical analy sis indicated that the difference was significant (P＜0.05). Conclusion   RNA interference targeting HIF-1αgene could suppress the proliferation of human tongue cancer cells effectively. HIF-1αgene could be researched as a new therapeutic target of tongue cancer in the future.

Key words: tongue neoplasms, cell line, tumor, RNA interference, hypoxia-inducible factor 1, alpha subunit, reverse transcriptase polymerase chain reaction, 印迹法