The Effects And Mechanism Of DiallylTrisulnde(DATS) Derivatives On Human Prostate Cancer PC-3 Cells

The aim of the present study is looking for the compounds can be able to inhibit tumor cell proliferationa efficiently from a serial of novel DATS derivatives which self-synthisized by own lab early, and the possible mechanism of action was discussed, which will be helpful for developing new anticancer agents with Self-dominated Intellectual Property Right.Androgen-independent prostate cancer PC-3 cells is chosen, and DATS derivatives including dibutenyl trisulfide(DBTS), bis(2-methylallyl) trisulfide (2-M-DATS). dipentenyl trisulfide(DPTS), bis(3-methylbut-2-enyl) trisulfide (3-M-DBTS) and dihexenyl trisulfide(DHTS) depressant effect of DATS and its derivatives on PC-3 is assayed by MTT Typical apoptotic nuclei were shown by Hochest33342 fluorescein staining and fluorescence microscope with 80μM concentrations of DATS and its derivatives for 24 h.. Introducing DNA Ladder experiment to validate that DATS and its derivatives can induce the Apoptosis of PC-3, meanwhile, Cell apoptosis rate was detected by flow cytometry with Annexin-V/PI staining. the mechanism of anti-cancer is also researched based on dibutenyl trisulfide(DBTS), bis(2-methylallyl) trisulfide (2-M-DATS), dipentenyl trisulfide(DPTS). Cell cycle is analyzed following PI staining by flow cytometry. Caspase-3 and Influence of proteinum relate to apoptosis is detected by Western bloting.PC-3 treated by different concentration of DATS and its derivatives after 24h and 48h,every compound can inhibit proliferation of PC-3 in vitro and the inhibition in significant does-and time-dependent manner. Bis(2-methylallyl) trisulfide (2-M-DATS) has the best effect of inhibiting proliferation, dibutenyl trisulfide(DBTS), dipentenyl trisulfide(DPTS) has fair inhibited effect compared to DATS, but, the inhibited effect of bis(3-methylbut-2-enyl) trisulfide (3-M-DBTS), dihexenyl trisulfide(DHTS) is weaker than it of DATS.After treation of DATS and its derivatives, some classical morphologic change of apoptosis cells are observed such as karyopyknosis, caryotin and karyorrhexis by Hochest33342 fluorescein stain and fluorescence microscope. According to DNA agarose gel electrophoresis, it can be observed the obvious step-form DNA Ladder which means cell apoptosis occurs and at the advanced stage of apoptosis. The result of Annexin V/PI staining demonstrated that apoptosis rate increased remarkably compared to control after treatment with DATS and its derivatives, and this result also confirmed that DATS and its derivatives can definitely induce apoptosis of PC-3. The result of FCM revealed that DATS and dibutenyl trisulfide(DBTS), bis(2-methylallyl) trisulfide (2-M-DATS), dipentenyl trisulfide(DPTS) arrested PC-3 cells at G2/M phase, and the percentage of G0/G1 phase cells decreased, the percentage of G2/M phase cells increased, and the percentage of S phase cells no significanted varied. According to the Western bloting assay, the result indicated that decomposition of caspase-3 can be stimulated, the express of Bcl-2 protein can be inhibited and up-regulation the express of Bax protein.In summary, DATS and its derivatives can inhibit prolifration of PC-3, induce apoptosis and arrest cell cycle. The mechanism probably is related to the increasing of cutting of caspase-3, down regulation of anti-apoptosis factor proteins Bcl-2 expression and the up-regulation of pro-apoptotic proteins Bax expression.