目的研究热疗对Tea8113细胞的侵袭能力及侵袭转移相关蛋白——基质金属蛋白酶-13(MMP-13)及钙离子结合蛋白S100A4(S100A4)表达的影响。方法Tea8113细胞分别经43℃加热0、40、80、120min及37、41、43、45℃加热80min,用体外侵袭实验评价各组细胞的侵袭能力;Tca8113细胞爬片经43℃分别加热0、40、80、120rain,用免疫细胞化学染色法检测各组细胞MMP-13及S100A4的表达变化;Tea8113细胞分别经43℃加热0、40、80、120min及37、41、43、45℃加热80min,Westernblot法检测各组细胞MMP一13及S100A4的表达变化。结果随加热时间延长或加热温度升高,Tea8113细胞侵袭率(即侵袭细胞数占全部细胞的百分比)依次递减,除43℃加热40、80min组之间及41、43℃加热80min组之间的差异无统计学意义(p〉0.05)外,其余各组问的差异均有统计学意义CP(0.05);随着加热时间延长,Tca8113细胞的细胞浆内MMP-13及S100A4的表达强度呈现递减趋势;随着加热时间延长或加热温度升高,Tca8113细胞的MMP-13及S100A4含量逐渐降低,除43℃加热40、80min组之间及41、43℃加热80min组之间外,其余各组间的差异均有统计学意义(P〈0.05)。结论热疗能抑制Tca8113细胞的侵袭能力,其机制可能与热疗抑制了细胞内MMP-13及S100A4的表达有关。
Objective To study the effect of hyperthermia on anti-invasion of Tea8113 and the expression change of matrix metalloproteinase-13 (MMP-13) and calcium-binding protein S100A4 (S100A4). Methods Tca8113 cell pools were incubated at 43 ℃ for 0, 40, 80, 120min, respectively, and at 37, 41, 43, 45℃ respectively for 80 min. The effect of high temperatures on the invasion ability of Tca8113 was measured in vitro. The slides of cells were made and incubated at 43℃ for 0, 40, 80, 120 min, respectively. Immunocytoehemical method was employed for detecting the expression change of MMP-13 and S100A4 protein. Tca8113 cells were incubated at 43℃ for 0, 40, 80, 120 min respectively and at 37, 41, 43, 45℃ respectively for 80 min. Western blot method was conducted for detecting the expressionchange of MMP-13 and S100A4 protein. Results As incubating time at higher temperature lasted, the proportion of the cells with invasion ability decreased. Except groups of 40 min and 80 min at 43℃ and 41, 43℃for 80min, the rest groups show significant statistic differences(P〈0.05). The expression intensity of MMP-13 and SIOOA4 proteins in Tea8113 cells would decrease as incubating time at higher temperature lasted. The content of MMP-13 and S100A4 proteins would decrease as incubating time at higher temperature lasted or incubating temperature increased.Except the groups of 40, 80 min at 43 ℃ and gl, 43℃ for 80 min, statistic differences were identified (P〈0.05). Conclusion The invasion of Tca8113 could be inhibited by hyperthermia. The mechanism of this effect may be due to protein expression inhibition of MMP-13 and S100A4.