环靶明联合吉非替尼对前列腺癌细胞株DU145增殖和侵袭的影响

目的：检测经环靶明与吉非替尼联合作用后前列腺癌DU145细胞生物学行为的变化，并探讨相关作用机制。方法前列腺癌DU145细胞经不同药物分组处理24 h后，采用MTT法和Transwell小室法检测细胞增殖及侵袭能力的改变；Western blot检测细胞中SHH、PTCH和Gli-1蛋白表达水平的变化。结果联合用药组细胞吸光度值(0.3842±0.0518)明显低于环靶明组(0.6839±0.0989)(t=4.6487，P=0.001)和吉非替尼组(0.7726±0.1474)(t=6.0245，P<0.01)，其平均穿膜细胞数(28.3333±3.3863)少于环靶明组(39.5000±5.6480，t=3.0677，P=0.036)和吉非替尼组(41.1667±6.3377)，(t=3.5256，P=0.013)，各实验中药物处理组与空白对照组间的差异均有统计学意义(P<0.01)；环靶明组及联合用药组各通路蛋白的表达均低于空白对照组(P<0.01)，吉非替尼组仅PTCH蛋白表达(0.7531±0.1287)低于空白对照组(1.1112±0.1157)(t=4.1495，P=0.019)，联合用药组的PTCH蛋白表达(0.3601±0.0900)低于环靶明组(0.6767±0.0815)(t=3.6686，P=0.038)和吉非替尼组(0.7531±0.1287)(t=4.5539，P=0.011)。结论环靶明联合吉非替尼较单独用药明显增强了对前列腺癌细胞增殖和侵袭的抑制作用，其机制可能与吉非替尼协同环靶明下调Hedgehog通路的PTCH蛋白表达有关。

Effects of combined application of cyclopamine and gefitinib on the proliferation and invasion of prostate cancer cell line DU145

Objective To detect the changes of biological behavior of DU145 cells after treatment with com-bined application of cyclopamine (CP) and gefitinib (G), and to explore the corresponding mechanism. Methods After DU145 cells were treated with different drugs for 24 h, the cell proliferation and invasion abilities were detected respec-tively by MTT method and transwell assay. The protein expressions of Sonic Hedgehog (SHH), Patched (PTCH) and GLI family zinc finger 1 (Gli-1) were detected by Western blot. Results The absorbance value of cells in combined drug group GC of (0.384 2±0.051 8) was significantly lower than that in group CP of (0.683 9±0.098 9) (t=4.6 487, P=0.001) and group G of (0.772 6 ± 0.147 4) (t=6.024 5, P<0.01). The average invasive cells of group GC of (28.333 3 ± 3.3863) was significantly less than that of group CP of (39.500 0 ± 5.648 0) (t=3.067 7, P=0.036) and group G of (41.166 7±6.337 7) (t=3.5 256, P=0.013). All the differences between drug groups and blank control groups GB were sta-tistically significant (P<0.01). All the expressions of SHH, PTCH and Gli-1 proteins in group CP and GC were signifi-cantly lower than those in GB (P<0.01). In group G only PTCH expression (0.753 1 ± 0.128 7) was significantly lower than that of group GB of (1.111 2±0.115 7), t=4.149 5, P=0.019;PTCH expression of GC (0.360 1±0.090 0) was lower than that of group CP of (0.676 7±0.081 5) (t=3.668 6, P=0.038) and group G of (0.753 1±0.128 7) (t=4.553 9, P=0.011). Conclusion Compared with single drug therapy, combined application of cyclopamine and gefitinib is more effective for inhibiting the proliferation and invasion abilities of prostate cancer cells, which might be related with both synergisti-cally down regulating expressions of PTCH in the Sonic hedgehog-Patched-Gli (Shh-Ptch-Gli) pathway.