目的:
    研究 HSP 的表达与乳腺癌细胞侵袭转移能力的相关性.
方法:
    以高转移人乳腺癌细胞株 MDA-MB-435s 和 MDA-MB-231为研究对象.MTT 法检测 HSP90 抑制剂 GA 对两株乳腺癌细胞株的增殖抑制作用,求出两株细胞的 IC50,采用半量 IC50检测 HSP90 抑制剂GA 对肿瘤细胞粘附、侵袭转移能力的影响;MTT 法检测细胞粘附Matrigel 能力;Transwell 小室进行人工重组基底膜侵袭和运动实验;RT-PCR 检测两株细胞 HSP90、HSP70 和 HSP27mRNA 水平的表达差异;Western-blot 检测两株细胞 HSP90 、HSP70 和 HSP27 蛋白质水平的表达差异.
结 果 :
    GA 对 高 转 移 人 乳 腺 癌 细 胞 株 MDA-MB-435s 和MDA-MB-231 均有细胞增殖抑制作用,且随药物浓度增大和时间延长而增强,其中对 MDA-MB-435s 细胞的 IC50为 32.08nM,半量浓度为16.0nM, 对 MDA-MB-231 细胞的 IC50 为 54.4nM,半量浓度为 27.2nM. MDA-MB-435s 和 MDA-MB-231 细胞侵袭转移能力没有显著差异(P﹥0.05).GA 可使两株乳腺癌细胞株对基底膜粘附能力下降,具有显著性(P<0.01).
    MDA-MB-435s 细胞对照组和实验组侵袭迁移实验穿膜细胞数分别为:113.2±6.42,41.2±9.04;132.6±6.58,57.8±9.42,
    MDA-MB-231 细胞对照组和实验组侵袭迁移实验穿膜细胞数分别为:120.4±12.64,101.0±5.24;140.4±10.50,118.0±11.73.
    MDA-MB-435s细胞实验组的侵袭迁移能力低于对照组,差异有非常显著性(P<0.01);
    MDA-MB-231 细胞实验组的侵袭迁移能力明显低于对照组,差异有显著性(P<0.01).
Objectives:
    To investigate the correlation between expression of HSP and invasive and metastatic potential in human breast cancer cell.
Methods:
    The highly metastatic human breast cancer cell lines MDA-MB-435s and MDA-MB-231 were used in this study. The effects of HSP90 inhibitor GA on the proliferation of breast cancer cell were explored by MTT colorimetry for obtaining their IC50. Adhesion of MDA-MB-435s and MDA-MB-231 to Matrigel was measured with MTT assay. The invasive and metastatic abilities of human breast cancer lines MDA-MB-231 and MDA-MB-435s in vitro were explored by transwell chamber. The expression of HSP27、HSP70、HSP90 in MDA-MB-231 and MDA-MB-435s cells was detected by RT-PCR and Western-blotting.
Results:
    GA showed a time/dose-dependent growth inhibition effect on MDA-MB-435s and MDA-MB-231 cells, IC50 of GA to MDA-MB-435s cell was 32.08nM and IC50 of GA to MDA-MB-231 cell was 54.4nM,The dose of GA used to treat the breast cancer cell lines were half of its IC50,i.e,16nM and 27.2nM respectively. Before treatment with GA,the ability of invasion and metastasis in MDA-MB-435s and MDA-MB-231 cells had no difference(P﹥0.05). After treatment with GA the ability of invasion and metastasis in MDA-MB-435s and MDA-MB-231 cells were significantly difference. (P<0.01). After treatment with GA the ability of invasion and metastasis in MDA-MB-435s lower than MDA-MB-231 cells (P<0.01).Compared with untreated group,adhesion of MDA-MB-435s and MDA-MB-231 were lower.The invaded cell numbers in invasive and metastatic experiments of MDA-MB-435s cell lines untreated or treated with GA were 113.2±6.42,41.2±9.04;132.6±6.58,57.8±9.42 respectively, the invasive and metastatic abilities of treated group were lower than untreated group significantly(P<0.01).The invaded cell numbers in invasive and metastatic experiments of MDA-MB-231 cell lines untreated or treated with GA were 120.4 ± 12.64 、 101.0 ± 5.24;140.4 ± 10.50 、 118.0 ± 11.73 respectively,the invasive and metastatic abilities of treated group were lower than untreated group significantly(P<0.05). The protein expression level of HSP90 in MDA-MB-435s was higher than MDA-MB-231 cell (P<0.01). The mRNA expression level of HSP90αin MDA-MB-435s was higher than MDA-MB-231 cell (P<0.01). The mRNA expression level of HSP90βhas no signifcant difference(P﹥0.05). The protein and mRNA expression level of HSP27 in MDA-MB-231 was higher than MDA-MB-435s cell (P<0.01). The protein expression level of HSP70 in MDA-MB-435s and MDA-MB-231 cells had no significant difference. (P ﹥ 0.05), The mRNA expression level of HSP70 in MDA-MB-231 was higher than MDA-MB-435s cell (P<0.05). Conclusion: The invasive and metastatic abilities of MDA-MB-435s and MDA-MB-231 cells were inhibited by HSP90 inhibitor GA.Consistent with HSP expression patterns in these two cell lines, MDA-MB-435s were inhibited more by GA than MDA-MB-231.Results had shown high expression of HSP90 in MDA-MB-435s and HSP27 in MDA-MB-231.It demonstrated that the HSP expression patterns were associated with the invasive and metastatic abilities of breast cancer lines.

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