疏肝益肾方对三苯氧胺耐药细胞株雌激素受体α和人表皮生长因子受体2表达的影响

Effect of Shugan Yishen recipe on expressions of estrogens receptor α and human epidermal growth factor receptor-2 in tamoxifen-resistant cell line

目的研究疏肝益肾方对三苯氧胺耐药乳腺癌细胞株LCC9细胞雌激素受体α（ERα）和人表皮生长因子受体2（HER2）表达的影响。方法采用40只健康Sprague Dawley大鼠分别制备疏肝益肾方、三苯氧胺、疏肝益肾方联合三苯氧胺及0.9%氯化钠溶液的含药血清（n=10），将LCC9细胞分为疏肝益肾方组、三苯氧胺组、联合组及对照组，分别以上述含药血清处理48 h。采用免疫细胞化学及免疫荧光法检测4种血清对LCC9细胞ERα、HER2蛋白表达的影响；免疫共沉淀法观察LCC9细胞中ERα及HER2之间的相互作用。结果ERα和HER2阳性细胞率三苯氧胺组与对照组比较差异无统计学意义（P＞0.05），疏肝益肾方组和联合组ERα阳性细胞率高于、HER2阳性细胞率低于三苯氧胺组和对照组［（79±7）%、（93±16）%比（63±19）%、（59±14）%；（83±12）%、（74±63）%比（85±51）%、（91±3）%］，联合组ERα阳性细胞率高于、HER2阳性细胞率低于疏肝益肾方组，差异有统计学意义（P＜0.05）；ERα和HER2表达水平三苯氧胺组与对照组比较差异无统计学意义（P＞0.05），疏肝益肾方组和联合组ERα表达水平高于、HER2表达水平低于三苯氧胺组和对照组［（1.07±0.10）、（1.18±0.12）比（0.89±0.10）、（1.04±0.20）；（1.06±0.99）、（0.96±0.73）比（1.24±0.17）、（1.34±0.59）］（P＜0.05），联合组ERα表达水平高于、HER2表达水平低于疏肝益肾方组（P＜0.05）。免疫共沉淀显示ERα与HER2之间无直接相互作用。结论疏肝益肾方可上调三苯氧胺耐药乳腺癌细胞株LCC9细胞中ERα的表达、下调HER2的表达，耐药细胞株中ERα与HER2之间无直接相互作用。

ObjectiveTo explore the expressions of estrogens receptor α （ERα） and human epidermal growth factor receptor-2 （HER2） in tamoxifen （TAM）-resistant breast cancer cell line LCC9. MethodsMedicated serums of Sprague Dawley rats containing Shugan Yishen recipe, TAM, Shugan Yishen recipe ＋ TAM and 0.9% sodium chloride （each n=10） were prepared; LCC9 cells were divided into Shugan Yishen recipe group, TAM group, combined group and control group treated with above medicated serums for 48 h, respectively. The expressions of ERα and HER2 were measured by immunocytochemistry and immunofluorescence; the interaction of ERα and HER2 was detected by immune co-precipitation. ResultsThe positive expression rates of ERα and HER2 in TAM group were not significantly different from those in control group （P〉0.05）; in Shugan Yishen recipe group and combined group the positive expression rates of ERα were significantly higher, the positive expression rates of HER2 were significantly lower than those in TAM group and control group [（79±7）%, （93±16）% vs （63±19）%, （59±14）%; （83±12）%, （74±63）% vs （85±51）%, （91±3）%] （P〈0.05）; the positive expression rate of ERα was significantly higher, the positive expression rate of HER2 was significantly lower in combined group compared with those in Shugan Yishen recipe group （P〈0.05）. The expression levels of ERα and HER2 in TAM group were not significantly different from those in control group （P〉0.05）; in Shugan Yishen recipe group and combined group, the expression levels of ERα were significantly higher, the expression levels of HER2 were significantly lower than those in TAM group and control group [（1.07±0.10）, （1.18±0.12） vs （0.89±0.10）, （1.04±0.20）; （1.06±0.99）, （0.96±0.73） vs （1.24±0.17）, （1.34±0.59）]（P〈0.05）; the expression level of ERα was significantly higher, the expression level of HER2 was significantly lower in combined group compared with those in Shugan Yishen recipe group （P〈0.05）. No direct interaction of ERα and HER2 was shown. ConclusionShugan Yishen recipe can effectively up-regulate the expression ER alpha expression and down-regulate the expression of HER2 in TAM-resistant breast cancer cell line, but no direct interaction between ERα and HER2 was found.