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目的:探讨5-氮杂-2’-脱氧胞苷(5-aza-2’-deoxycytidine,5-aza-dC)联合曲古菌素A(trichostatin A,TSA)能否诱导性激素受体阴性的乳腺癌细胞系SKBR-3同时重新表达功能性雌激素受体α(estrogen receptoralpha,ERα)和雄激素受体(androgen receptor,AR),并进一步研究诱导前后SKBR-3细胞对乳腺癌内分泌治疗敏感性的变化。方法:分别采用RT-PCR法和免疫细胞化学法检测5-aza-dC联合TSA诱导前后SKBR-3细胞和阳性对照MCF-7细胞中ERα、AR、孕激素受体(progestone receptor,PR)、雌激素调节蛋白pS2与前列腺特异性抗原(prostate specific antigen,PSA)mRNA和蛋白的表达水平;采用CCK-8法检测各内分泌治疗药物对5-aza-dC联合TSA诱导后重新表达ERα和AR的乳腺癌SKBR-3细胞增殖能力的影响。结果:5-aza-dC联合TSA能够诱导激素受体阴性的乳腺癌SKBR-3细胞同时重新表达ERα和AR,它们的下游产物PR、pS2以及PSA也相应表达;诱导后SKBR-3细胞中ERα和AR的表达量明显低于在MCF-7细胞中的表达量(P<0.05)。5-aza-dC联合TSA能明显抑制SKBR-3细胞的增殖能力(P<0.05),加入雌激素类药物17β-雌二醇(17β-estradiol,E2)后细胞的增殖能力略有上升,但差异无统计学意义(P>0.05)。在加入E2的基础上,分别再加入雌激素拮抗剂他莫昔芬(tamoxifen,TAM)或孕激素醋酸甲地孕酮(megestrol acetate,MA)后,二者都能使肿瘤细胞的增殖能力明显下降(P<0.05);联合加入TAM和MA后,肿瘤细胞的增殖能力进一步明显下降(P<0.05)。结论:5-aza-dC联合TSA能够诱导乳腺癌细胞系SKBR-3同时恢复表达功能性的ERα和AR,抑制肿瘤细胞的生长,同时使SKBR-3细胞恢复了对激素的依赖性和内分泌治疗的敏感性。
Objective: To investigate whether 5-aza-2’-deoxycytidine (5-aza-dC) combined with trichostatin A (TSA) can induce hormone receptor negative The breast cancer cell line SKBR-3 simultaneously re-expressed functional estrogen receptor (ERα) and androgen receptor (AR) and further studied the sensitivity of SKBR-3 cells to endocrine therapy of breast cancer before and after induction The change. Methods: The expressions of ERα, AR, progestone receptor (PR) and the mRNA expressions of ERα, AR in SKBR-3 cells before and after 5-aza-dC combined with TSA were detected by RT-PCR and immunocytochemistry The expressions of estrogen-regulated protein pS2 and prostate specific antigen (PSA) mRNA and protein were detected by immunohistochemistry. The expression of ERα and AR in 5-aza-dC combined with TSA was detected by CCK-8 assay Effect of Breast Cancer SKBR-3 Cell Proliferation. Results: 5-aza-dC combined with TSA could induce estrogen-receptor negative breast cancer SK ER-3 and AR re-expressing ER, AR and their downstream products PR, pS2 and PSA respectively; And AR were significantly lower than those in MCF-7 cells (P <0.05). The proliferation of SKBR-3 cells was significantly inhibited by 5-aza-dC combined with TSA (P <0.05). Proliferation of cells increased slightly with the addition of 17β-estradiol (E2) The difference was not statistically significant (P> 0.05). On the basis of adding E2, after adding estrogen antagonist tamoxifen (TAM) or progesterone megestrol acetate (MA) respectively, both of them can make tumor cell proliferate obviously (P <0.05). After adding TAM and MA, the proliferation of tumor cells was significantly decreased (P <0.05). CONCLUSIONS: 5-aza-dC combined with TSA can induce breast cancer cell line SKBR-3 to simultaneously restore functional ERα and AR expression, inhibit the growth of tumor cells and restore hormone-dependent and endocrine therapy of SKBR-3 cells Sensitivity.