Aqueous Extract Of Huang-lian Induces Apoptosis In Lung Cancer Cells Via P53- Mediated Mitochondrial Apoptosis

Objective: The proliferation of human lung cancer A549 cells and apoptosis pathway effected by Huang Lian extract was studied. Methods: The study was divided into test group(10μg/mL,20μg/mL,40μg/mL,60μg/mL) and control group, to detected the MTT value at 12, 24 and 48 h. The apoptosis status effected by aqueous extract of Huanglian was detected by TUNEL method. The expression level of P53, Bax, Bcl-xl and Bcl-2 effected by aqueous extract of Huanglian(concentration of 40 μ g/mL) in A549 cells was detected by Real-time PCR and Western blot. The Caspase-3 and the cleaved Caspase-3 was detected by Western blot. After to interved P53, inhibitors P53 was used to dected Bax, Bcl-xl and Bcl-2 changes in mRNA and protein expression. Results: The maximum inhibition rate and apoptosis appeared at an aqueous extract of Huanglian concentration of 40μg/mL on the 48 h of treatment. The mRNA and protein expression levels of p53, Bcl-2 and Bcl-xL decreased as the time increased. However, Bax expression increased with increasing time. The cleavaged caspase-3 increased as the time increased. The p53 was inhibited by PFT-α(p53 inhibitor). The Bcl-2 and Bcl-xL did not increased as the time increased. But, the Bax was increasted. Conclusion: Huang-lian acts by regulating the Bcl-2 family protein-mediated mitochondrial pathway via p53 and provide evidence that Huang-lian deserves further investigation as a natural agent for treating and preventing cancer.