| Objective1. Examine the changes of Kiss-1mRNA and protein expression in Human colorectal cancer SW480 cells which is the most in Kiss-1 protein and Kiss-1 mRNA expressed, after the cells were exposed to X-ray irradiation at 24 and 48 hours later, and find the relationship between the changes of Kiss-1 gene expression and the characteristic cytobiology of proliferation, apoptosis, migration and invasion, after the human colorectal carcinoma irradiated by the X-ray;2. Investigate the effects of radiation on metastasis suppressor gene Kiss-1 expression of rectal carcinoma.Methods:1 Detect the location where kiss-1 expressed in SW480 cells by immunohisto -chemistry. Compare the expression of Kiss-1mRNA and Kiss-1 protein in human colorectal carcinoma cells among SW480, HCT116 and SW1116;2 Irradiate the SW480 with a single different X-ray (0, 2, 4, 6, 8Gy) respectively, the Kiss-1 expression was examined by SYBR Green quantitative real-time PCR and immunohistochemistry after 24 and 48 hours;3 Colorectal cancer cell line SW480 was irradiated at different does (0, 2, 4, 6, 8Gy). After 24 and 48h, the cell immunohistochemistry was used to find out the effect of X-ray to the Kiss-1 protein expression of SW480; colony formation assay was used to verify the proliferation of SW480; the flow cytometry-Annexin-V/PI assays was used to observe the changing of the apoptosis rate; the Transwell migration model was established by the Transwell chamber of 8um diameter, was used to observed the migration of SW480;the Transwell invasion model was established by Transwell chamber of 8 um diameter and ECM as the basement membrane, was used to observe the invasion of SW480;4 The expressions of Kiss-1 were examined immunohistochemically with SP method in 29 specimens of rectal carcinoma tissues before radiation and after radiation. Results1 In colorectal cancer SW480 cells, Kiss-1 is mainly located in the endochylema of cells. The expression of Kiss-1 protein and Kiss-1mRNA in SW480 cells is higher than in S41116 and HCT116;2 In SW480 cells,the expression of kiss-1 at 24 hours after irradiation increase significantly, but doesn't change as the dose increasing from 10Gy to 20Gy; at 48 hours after irradiation, the KISS-1 expression in cells irradiated with 2Gy, 10Gy and 20Gy, are much higher than the control, and almost the same as each other, no matter how much the dose. Compare with the expression at 24 hours after irradiation, the KISS-1 expressions at 48 hours don't change much in cells irradiated with 10Gy and 20Gy, but increase in those irradiated with 2Gy;3 Compare with the no irradiation group, 24h or 48h after radiation, the proliferation of SW480 was increased 48h after radiation, but there was no change 24h after radiation; the apoptosis of SW480 were increased for the 4, 6, 8Gy 48h after radiation, there was no significant difference for 2Gy groups; in the migration and invasion assay, the migration and invasion of SW480 was significantly increased after irradiation including 4Gy and 8Gy group;4 Before and after accepeting general DT50GY/25f/5 week regimen of pelvic radiotherapy on rectal cancer, the expressions of Kiss-1 gene in rectal carcinoma did not change obviously.Conclusion1 The Kiss-1 gene expression of colorectal cancer cells can be increased by appropriate intensity-modulated X-ray radiation dose. 2 X-ray radiation inhibit tumor cell proliferation and promote apoptosis of tumor cells may be associated with increased Kiss-1 gene expression; Kiss-1 gene may be a potential assessment of colorectal cancer recurrence after radiotherapy biological indicators;3 X-ray irradiation of colorectal cancer cell invasion, migration may be related to Kiss-1 gene expression is not related, Kiss-1 gene is not yet used to evaluate tumor metastasis of colorectal cancer risk after radiotherapy of independent biological indicators. |
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