华蟾素注射液对乳腺癌MDA-MB-231细胞E-cad、N-cad表达的影响

Effect of Huachansu Injection on E-cad and N-cad in Breast Cancer Cell Line MDA-MB-231

目的观察华蟾素注射液对乳腺癌MDA-MB-231细胞E-cad、N-cad表达的影响,探讨华蟾素注射液对乳腺癌细胞上皮间质转化(EMT)的影响,探讨华蟾素注射液抑制乳腺癌转移机制。方法采用四氮唑蓝(MTT)比色法观察高、中、低剂量华蟾素注射液(100mg/L、25mg/L、6.25mg/L)对MDA-MB-231细胞增殖的影响;实时荧光定量PCR(q RT-PCR)检测法检测MDA-MB-231细胞中E-cad/N-cad m RNA表达水平;蛋白印迹法(Westernblot)检测MDA-MB-231细胞中E-cad/Ncad蛋白表达水平。结果高浓度组华蟾素注射液48h对MDA-MB-231细胞抑制率较中、低剂量组明显(54.54%比50.20%、37.80%),差异有统计学意义(P<0.05),q RT-PCR实验结果显示空白对照组、华蟾素注射液高剂量组、中剂量组、低剂量组E-cad相对表达量依次为(1.05±0.04)%、(8.77±1.54)%、(3.74±1.56)%、(1.64±0.30)%,N-cad相对表达量依次为(1.18±0.19)%、(0.34±0.19)%、(0.79±0.20)%、(0.84±0.07)%。华蟾素可以上调E-cad表达,下调N-cad表达,与空白对照组比较差异有统计学意义(P<0.01)。Westernblot结果显示华蟾素注射液能降低N-cad蛋白的表达,上调E-cad蛋白的表达。结论华蟾素能通过上调E-cad表达和下调N-cad表达,抑制MDA-MB-231细胞的上皮间质转化(EMT)转变,从而降低乳腺癌MDA-MB-231细胞的转移能力。

Objective To investigate the effect of huachansu injection on the expressions of E-cad and N-cad in breast cancer cell line MDA-MB-231, in order to explore the inhibiting effect of huachansu injection on the metastasis of breast cancer. Methods MTT assay was applied to observe the proliferation of MDA-MB-231 cells after treatment with high, medium and low doses of huachansu injection（100mg/L, 25mg/L and 6.25mg/L）. The expressions of E-cad and N-cad m RNA and protein in test cells were detected by q RT-PCR and western blot, respectively. Results The high dose group of huachansu injection had a higher cell inhibition effect at 48 hours compared with the medium dose group and low dose group（54.54% vs 50.20%, 37.80%, P〈0.05）. q RT-PCR results showed that the expressions of E-cad m RNA in high, medium and low doses of huachansu injection groups were up-regulated and the expression of N-cad m RNA was up-regulated compared with those in blank control group [E-cad：（8.77 ±1.54）%,（3.74 ±1.56）%,（1.64 ±0.30）% vs（1.05 ±0.04）%, P 0.01; N-cad：（0.34 ±0.19）%,（0.79±0.20）%,（0.84±0.07）% vs（1.18±0.19）%, P〈0.01] and the similar results in E-cad and N-cad proteins were observed by western blot（P〈0.05）. Conclusion Huachansu injection can inhibit the EMT transition of MDA-MB-231 cells through down-regulating N-cad and up-regulating E-cad, so as to decreasing the migration and invasive ability of the cancer cells.