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目的观察不同温度下吉西他滨(GEM)热化疗对膀胱癌BIU-87细胞株侵袭能力的影响,并探讨其可能的机制。方法将处于对数生长期的BIU-87细胞分为4组,A组为对照组,B组为热化疗1组[(38±0.2)℃],C组为热化疗2组[(40±0.2)℃],D组为热化疗3组[(43±0.2)℃],GEM的工作浓度为1μg/ml。用Transwell小室实验检测各组细胞的侵袭能力;用RT-PCR检测MMP-9m RNA的表达情况;用ELISA法检测各组细胞的MMP-9蛋白的表达情况;用EMSA检测各组细胞NF-κB的活性。结果 Transwell小室实验显示相对于A组,B、C、D组对BIU-87细胞的穿透能力均可产生抑制作用,热化疗组与对照组之间的差异均有统计学意义(P<0.05),其中D组的抑制作用最为明显。ELISA检测结果显示与对照组相比,实验组各组均能抑制MMP-9蛋白的表达,热化疗组与对照组之间的差异有统计学意义(P<0.05),其中D组的抑制最为明显。EMSA实验结果显示实验组NF-κB的活性明显降低,与对照组之间的差异有统计学意义(P<0.05)。结论热化疗能够抑制BIU-87细胞的侵袭,并且随着温度的升高抑制作用越明显,其机制可能是通过抑制NF-κB的活性,降低MMP-9蛋白的表达来发挥作用的。
Objective To investigate the effect of gemcitabine (GEM) thermotherapy on the invasiveness of bladder cancer cell line BIU-87 at different temperatures and to explore its possible mechanism. Methods BIU-87 cells in logarithmic growth phase were divided into 4 groups, group A was control group, group B was thermochemotherapy group [(38 ± 0.2) ℃], group C was thermochemotherapy group (40 ± 0.2) ° C], group D was 3 groups [(43 ± 0.2) ° C], and the working concentration of GEM was 1 μg / ml. The invasion ability of each group of cells was detected by Transwell chamber assay. The expression of MMP-9mRNA was detected by RT-PCR. The expression of MMP-9 protein was detected by ELISA. The expression of NF-κB Activity. Results Transwell chamber experiments showed that BIU-87 cells could inhibit the penetration ability of BIU-87 cells compared with those of A, B, C and D groups, and there was significant difference between thermo-chemotherapy group and control group (P <0.05 ), Of which D group the most obvious inhibition. The results of ELISA showed that compared with the control group, the experimental group inhibited the expression of MMP-9 protein, the difference between the thermochemotherapy group and the control group was statistically significant (P <0.05), of which the inhibition in group D was the most obvious. The results of EMSA showed that the activity of NF-κB in experimental group was significantly lower than that in control group (P <0.05). Conclusions Thermochemotherapy can inhibit the invasion of BIU-87 cells, and the more obvious the inhibitory effect with the increase of temperature, the mechanism may be through inhibiting the activity of NF-κB and reducing the expression of MMP-9 protein.