Abstract:

Objective To evaluate the effects of brevilin on HeLa cell proliferation，apoptosis，ROS production and mitochondrial dysfunction and its related mechanisms. Methods HeLa cell line was used as the study object. MTT method，flow cytometry，DCFH ⁃DA probe and mitochondrial swelling test were used to detect the effects of brevilin on HeLa cell proliferation，apoptosis，ROS production and mitochondrial function. Western blot was used to detect the specific protein expressions of mitochondrial dysfunction and PINK1/Parkin pathway. We constructed a small interfering RNA（siRNA）mediated low PINK1 expression cell line to verify the mechanism of brevilin. Results Brevilin inhibited cell viability，induced apoptosis and promoted the production of ROS in a dose⁃ dependent manner（P < 0.01）. Moreover，with the increase of brevilin concentration，the mitochondrial swelling of HeLa cells increased gradually（P < 0.001）. The ration of Bax/Bcl⁃2，Cyt C and cleaved caspase⁃3 upregulated in HeLa cells with the increasing concentration of brevilin（P < 0.01）. At the same time，the release of CytoCyt C were significantly enhanced（P < 0.01）and the level of Mito Cyt C clearly decreased（P < 0.01）after treatment with brevilin. Compared with the control group，the expression of PINK1 and Parkin protein in brevilin treated group was significantly lower（P < 0.01）. Further functional evaluation showed that PINK1 gene knockdown produced similar in vitro activity results as brevilin treatment，such as inhibiting HeLa cell activity，increasing the percentage of apoptotic cells and promoting ROS production. Conclusion Brevilin treatment activates the PINK1/Parkin pathway，and thereby inhibits HeLa cell viability，increases the percentage of apoptotic cells and promotes ROS production.

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