Effects Of TMPyP4-PDT On Human Cervical Cancer Hela、siha Cells And The Expression Of C-myc、hTERC

Cervical cancer is one of the most common cancer in women in the worldwide,where its incidence and mortality remain high in female cancer. The treatment depends on the range of lesions, clinical staging, patient’s age and fertility requirements,emphasis on individualized treatment. At present, surgery, chemotherapy, radiotherapy is still the traditional treatment of cervical cancer.Photodynamic therapy is a kind of new auxiliary tumor treatment that has rapid development in recent years, its main principle is to use the endogenous or exogenous photosensitizer selective absorption and retention in the tissue of the lesion, then under specific wavelength of laser irradiation, produced many cell toxic reactive oxygen species to achieve the aim of the cell or tissue death.5,10,15,20-tetra-(N-methyl-4-pyridyl)porphyrin,a four valence cationic porphyrin compound,which is a kind of synthetic new water-soluble photosensitizer, can be used to induce G-four conjoined formation of small molecular ligands and inhibit the activity of telomerase. This research will investigate the effect of photodynamic therapy mediated by meso-5,10,15,20-Tetrakis-(N-methyl-4-pyridyl) porphine (TMPyP4) on human cervical cancer hela、siha cells in vitro and the c-myc, hTERC gene expression in hela cell, discuss the possible mechanism of the action.PART1Effects of Photodynamic therapy with TMPyP4on the survival of human cervical cancer cells hela and siha in vitroObjective:To investigate the effects of TMPyP4-PDT on the survival of human cervical cancer cells hela and siha in vitro.Methods:Human cervical cancer cells hela and siha were divided into TMPyP4 group,PDT group,TMPyP4-PDT group and control group.The proliferative activity of hela and siha cells was assessed by CCK8assay.Results:TMPyP4group,PDT group,and control group has no inhibitory effect on the cells.Under the same drug concentration, with the strengthening of light energy, the inhibition rate of cell gradually strengthened in the TMPyP4-PDT group(P<0.05).Under the same energy, when the concentration of TMPyP4in0to50μmol/L, the inhibition rate increased with the increase of drug concentration.When the TMPyP4concentration reached50μmol/L,the inhibition rate went into the plateau50μmol/L TMPyP4、8J/cm2light dose were the optimal parameters for the killing of hela and siha cells in this experiment,and in this case,the50%inhibition concentration(IC50) of TMPyP4-PDT for hela and siha cells were8.158,14.693μmol/L respectively.(P<0.05).Conclusions:TMPyP4-PDT has distinctive inhibition effect on hela and siha cells,which correlated with concentration-dependent manner and energy-dependent manner.the sensitivities of the two cells to PDT are different.hela cells are more sensitive. PART2Effects of photodynamic therapy with TMPyP4on the expressions of c-my、hTERC in hela cellsObjective:To investigate the effects of TMPyP4-PDT on the expressions of c-myc、 hTERC in hela cells.Methods:Choose the laser dose4J/cm2.The modality of TMPyP4-PDT treated cells were observed with light microscope and apoptosis examined by flow cytometry under different concentration of TMPyP4. Choose the concentration of TMPyP420μmol/L,the expressing of c-myc mRNA、hTERC mRNA of the cell lines was tested by RT-PCR.Results:The necrosis and apoptosis cells were seen with light microscope.Annexin Ⅴ-FITC/PI double dye kits tests the rates of cell apoptosis and necrosis increased as TMPyP4concentration increased.The quantity of the expressing of c-myc mRNA、 hTERC mRNA was decreased as the extension of incubation time.Conclusions:TMPyP4-PDT has obvious killing effect on cervical cancer hela cells in vitro, and can induce its apoptosis, which correlated with concentration-dependent manner.The possible mechanism may be related with downregulating the expression of c-myc mRNA、hTERC mRNA.