[Impact of STAT6 signaling pathway blockade on apoptosis of human colon cancer cells].

OBJECTIVE

To investigate the relationship between the signal transducer and activator of transcription 6 (STAT6) and human colon cancer.

METHODS

Four STAT6-specific recombinant plasmid vectors, pshRNA-STAT6-1, 2, 3, and 4 were constructed and transfected into the cultured human colon cancer cells of the line HT-29. Seventy-two hours later RT-PCR was used to detect the mRNA expression of STAT6 and the apoptosis-related genes Bcl-2 and Bax, flow cytometry (FCM) was used to detect the protein expression of phopho-STAT6 (pSTAT6). HT-29 cells were inoculated into a plate and transfected with pshRNA-STAT6-1 or pshRNA-STAT6-4, and HT-29 cells without transfection were used as controls. Seventy-two hours later FCM was used to observe the cell apoptosis. Another HT-29 cells were inoculated into a plate and transfected with pshRNA-STAT6-1 or pshRNA-STAT6-4, or blank liposome as controls. Seventy-two hours later. Western blotting was used to detect the protein expression of Bcl-2 and Bax genes.

RESULTS

The p-STAT6 protein expression rate was 3.2% +/- 0.6% in the pshRNA-STAT6-1 group, significantly lower than that of the blank control group (18.2% +/- 0.9%, P < 0.01) with an inhibition rate of 82.4%, and was 7.9% +/- 0.4% in the pshRNA-STAT6-4 group, significantly lower than that in the blank control group too (P < 0.01) with an inhibition rate of 56.6%. And the p-STAT6 protein expression rates of the pshRNA-STAT6-2 and pshRNA-STAT6-3 groups were 16.6% +/- 0.5% and 17.1% +/- 0.7% respectively, both not significant different from that of the blank control group (both P > 0.05). The early cell apoptosis rates of the pshRNA-STAT6-1 and pshRNA-STAT6-4 groups were 13.0% and 8.8% respectively, both significantly higher than that of the blank control group (0.4%, both P < 0.05). The mRNA expression of Bcl-2 was significantly lower and the mRNA expression of Bax was significantly higher in the pshRNA-STAT6-1 and pshRNA-STAT6-4 groups than in the blank control and blank liposome groups (all P < 0.01). The protein expression patterns of Bcl-2 and Bax was consistent with that of their protein expression.

CONCLUSION

STAT6 signaling pathway inhibits the apoptosis of colon cancer cells by regulation of the Bcl-2 and Bax genes.