| Surgery, radiotherapy and chemotherapy are the three main traditional treatments of gynecological cancer. Using these methods will kill tumor cells while normal cells will also be inevitably damaged. These treatments often cause some serious complications, and sometimes have to be stopped. Some young unmarried or unpregnant patients strongly required the retention of their physiological and reproductive function and some elderly patients can not tolerate surgery, radiotherapy and chemotherapy, the others refuse surgical treatment and are resistant to chemotherapy, many of them are urgent for a gentle and effective treatment. Therefore photodynamic therapy (PDT) comes into being. Using PDT to treat malignant tumors has been achieved remarkable results and shown a good prospect. Different tumor has a sensitivity to PDT. The apoptosis induced by PDT does not reach to 100%. It is suggested that some protective factors against PDT exist in tumor cells. Some researchs show that the heat shock protein 70 (HSP70), heme oxygenase-1 (HO-1) and hypoxia inducible factor-1α(HIF-1α) in cancer cells can be activitied when the cells are in stress or exposed to some drugs in order to inhibit the apoptosis of cells. The aim of the research is to investigate the effects of 5-ALA-PDT on human gynecological carcinoma cells in vitro and expressions of HSP70,HO-1 and HIF-lα.PART 1 Effects of photodynamic therapy with 5-aminolevulinic acid on the survival of human gynecological carcinoma cells in vitroObjective:To investigate the effects of 5-ALA-PDT on the survival of human ovarian cancer cells SKOV3, cervical cancer cells Hela and endometrial cancer cells KLE.Methods:The 3 cell lines were divided into 5-ALA-PDT group,5-ALA group, PDT group and control group. We analyzed the effects of 5-aminolevulinic acid mediated photodynamic therapy (5-ALA-PDT) on SKOV3, Hela and KLE cells by using CCK-8 assay.Results:The inhibition rates of cells gradually increased with the increase of the 5-ALA concentration and laser irradiation dose (P<0.05). When the 5-ALA concentration and laser irradiation dose reached a certain level, the inhibition rate went into the plateau. The inhibition rates changed with time, they reached to peak in 24 hours after 5-ALA-PDT, then began to decline. The IC50 of SKOV3 was the highest of the 3 cell lines (P<0.05).Conclusions:5-ALA has photodynamic effects on SKOV3, Hela and KLE cells, there exists the phenomenons of concentration saturation and laser irradiation dose saturation; the inhibition rates change with time; the sensitivities of the 3 cell lines to PDT are different, the SKOV3 cell line is the least sensitive one.PART 2 Effects of photodynamic therapy with 5-aminolevulinic acid on the expressions of HSP70,HO-1 and HIF-la in SKOV3Objective: To investigate the effects of 5-ALA-PDT on the expressions of HSP70,HO-1 and HIF-la in SKOV3 cells.Methods: The SKOV3 cells were divided into 5-ALA-PDT group and control group. We analyzed the changes of the expressions of HSP70,HO-1 and HIF-1αin SKOV3 cells before and after 5-ALA-PDT by using Fluorescent real-time quantitative PCR.Results:The expressions levels of HSP70,HO-1 and HIF-1αin SKOV3 cells gradually increased over time (3h,12h,24h,48h,72h) after 5-ALA-PDT. The levels of the expressions of HSP70 and HIF-la significantly increased in 24hours after 5-ALA-PDT (P<0.05). The expression trends of HSP70,HO-1 and HIF-la were on the contrary of the trend of the inhibition rate.Conclusions:HSP70,HO-1 and HIF-1αmay be involved in the protection mechanism against stress induced by 5-ALA-PDT. They may be interrelated to the resistance of cancer cells to 5-ALA-PDT. |
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