The Role Of ID-1、Survivin And Caspase-3in ALA-PDT Induced Skin Squamous Cell Carcinoma A431Cells Apoptosis And Proliferation

Cutaneous squamous cell carcinoma (squamous cell carcinoma, SCC), is a kind of malignant tumor originated from cutaneous adnexal keratinocytes, occurs in the skin or mucosa, appendages, mainly from the squamous epithelium covering skin began to. Has a complex etiology, clinical misdiagnosis rate is high, malignant degree and invasive and destructive. Method in the treatment of cutaneous squamous cell carcinoma with operation excision, radiotherapy, photodynamic therapy and topical drug treatment. Photodynamic therapy (Photodynamic therapy, PDT) is also known as optical radiation therapy(Photoratiation, Therapy, PRT) or photochemical therapy (Photochemotherapy), is an effective method for treatment of benign development of malignant lesions, the surface of some part of the results, has been used in clinical. In recent years,5-ALA(5-aminolevulinic acid,5-aminolevulinic acid)mediated photo-dynamic therapy(5-ALA-PDT)treatment was successfully applied in non-melanoma skin cancer, but the mechanism remains to be investigated.Objective:1、To investigate the protein and mRNA expression of ID-1, Survivin and Caspase-3in human skin squamous cell carcinoma A431cells and its clinical significance;2、To investigate the inactivation effect of5-ALA-PDT on human skin squamous cell carcinoma A431cell;3、To observe the expression levels of ID-1, Survivin and Caspase-3in A431cells, explore the inner link and the possible relevance, and provide theoretical basis for clinical early diagnosis and treatment of squamous cell carcinoma.Method:1、Using Western blot and RT-PCR detect expression level of ID-1, Survivin and Caspase-3in human skin squamous cell carcinoma A431cells and keratinocytes (Hacat cells);2、In vitro experiment, A431cells and5-ALA in vitro cultivation after laser irradiation together, with MTT method to measure the survival rate, to explore the effect of culture time,5-ALA concentration and light dose of photodynamic effect mediated by; 3、The A431cells were divided into two groups (control group and experimental group), the previous conclusions on the basis of the experimental data, the control group without any treatment, detecting the ID-1, Survivin and Caspase-3protein of A431cells which after treatment with PDT(2h、4h、12h、24h、48h).Results:1、The protein and mRNA of ID-1and Survivin have a high expression in A431cells, and low expression in the Hacat cells with or without expression, expression was statistically significant difference (P<0.05); The Caspase-3protein and mRNA in the table on the contrary, the difference was statistically significant (P<0.05);2、At the same light dose and the concentration of5-ALA,5-ALA and A431cells co-cultured80min its the PDT killing effect mediated strongest;3、At the same light dose and incubation time,5-ALA concentration of3.2mmol/L, the mediated Photodynamic reached saturation;4、At the same incubation time and concentration of5-ALA, The light dose is80J/cm2, the inhibition of A431cells rate reached the maximum;5、The separate light and separate dosing have no effect on A431cells;6、The protein of ID-1and Survivin in5-ALA-PDT treated A431cells expression level with the extension of time has decreased, the difference was statistically significant (P<0.05);7、The protein of Caspase-3in PDT treated A431cells expression level with time extending upward trend, the difference was statistically significant (P<0.05).Conclusion:1、The high expression of ID-1and Survivin, low expression of Caspase-3may be related to the formation of skin squamous cell carcinoma;2、5-ALA-PDT can induce apoptosis in human squamous cell carcinoma A431cells;3、5-ALA-PDT-induced apoptosis in cutaneous squamous cell carcinoma A431cells, and its mechanism of action may directly or indirectly inhibit ID-1and Survivin expression and induced Caspase-3expression to promoting apoptosis.