Screen And Study The Mechanism Of Novel Porphyrins On The Anti-tumor Effect In Vitro

Objective: To observe the anti-tumor effect of newly synthesized metal porphyrins onthe hepatoma cells (HepG2)、gastric cancer cells (SGC-7901)、colon cancer cells (sw480)、nasopharyngeal carcinoma cells (CNE-1and CNE-2)、mouse monocyte macrophage(RAW264.7)、lung carcinoma cells(A549) and breast cancer cells(MCF-7), and screen thepotent drug to further study the mechanism of anti-tumor.Methods:1.To screen the anti-tumor effects of novel porphyrins:The proliferation ratiosof tumor cells were determined by the SRB and MTT method. The scavenging activities oftumor cells on hydroxyl free radicals and superoxide anion free radicals were detected bythe methods of phenanthroline-Fe2+oxidation and pyrogallol autoxidation. The contents ofNO in the cell culture supernate were detected by the Griess. The cell morphology wasobserved by the Giemsa staining.2.To research the mechanisms of anti-tumor drugs:Thechanges of mitochondrial membrane potential were detected with rhodamine123; Theapoptosis was assayed with flow cytometry; DNA integrity detection;The mRNAexpressions of C-myc mRNA were determined by the real-time PCR method;Thetelomerase activity was detected by the ELISA method.Results: All of the eight porphyrins had inhibitory effects on the growth of tumor cells.With the concentration of drugs increasing,the inhibition rates of cell growth wereincreased; the hydroxy free radicals and superoxide anion free radicals scavenging abilitywere correspondingly dropped; the contents of NO in the cell culture supernate wereelevated;the apoptotic cells were increased and the apoptotic HepG2cells weresignificantly increased by the Giemsa staining. Among eight drugs, Rup-03and Rup-04were the better for the above-mentioned inhibition effects. Rup-03had the better effects onHepG2、 CNE-1、CNE-2、sw480and RAW264.7cells, Rup-04had the better effects onSGC-7901cells.Within the increasing concentration(10-9~10-6mol/L) of Rup-03, thefluorescence intensity of mitochondrial membrane potential was decreased;the HepG2cells apoptosis rates were increased; agarose gel electrophoresis showed that DNA haddegraded; the relative expressions of C-myc mRNA were reduced and the telomerase activity was dropped. These results all indicated that the extent of apoptosis increased withthe drug concentration increasing.Conclusion: Egiht novel metal porphyrins all had killing effect on tumor cells, and thekilling effects have a drug-concentration-dependent.The best drug is Rup-03at10-9mol/Lto10-6mol/L. The anti-tumor mechanisms of novel porphyrins maybe produce the freeradicals via illumination to inhibit the expressions of C-myc gene and telomerase activitiesin tumor cells and induce apoptosis.