Ganguly Bhattacharjee Karabi, Bhattacharyya Mau, Halder Umesh Chandra, Jana Pradipta, Sinha Asru K
Sinha Institute of Medical Science and Technology, 288, Kendua Main Road, Garia, Kolkata, 700084, India.
Breast Cancer. 2014 Sep;21(5):605-13. doi: 10.1007/s12282-012-0422-6. Epub 2012 Nov 1.
Although progesterone receptor (PR) status, similarly to estrogen receptor status, is of prognostic importance in breast cancer, the involvement of the PR in breast cancer remains obscure. Studies were conducted to determine the function of the PR in neutrophils in the nitric oxide-induced synthesis of maspin, an anti-breast-cancer protein produced in nonmalignant mammary cells and in neutrophils in the circulation.
PR status was determined by immunohistochemistry. Maspin synthesis was determined by in-vitro translation of messenger RNA and quantified by enzyme-linked immunosorbent assay. Nitric oxide was determined by the methemoglobin method.
It was found that PR status in neutrophils was identical with that in malignant breast tissues. A Scatchard plot for progesterone binding to normal and PR-positive (PR+) neutrophils revealed that whereas normal neutrophils had 11.5 × 10(10) PR sites/cell with K d = 47.619 nM, PR+ neutrophils had 6.6 × 10(10) PR sites/cell with K d = 47.619 nM. The progesterone negative (PR-) neutrophils failed to bind to progesterone. Incubation of normal and PR+ neutrophils with 25 nM progesterone produced 1.317 μM NO and 2.329 nM maspin; the PR+ neutrophils produced 0.72 μM NO and 1.138 nM maspin. The PR- neutrophils failed to produce any NO or maspin in the presence of progesterone. Inhibition of progesterone-induced NO synthesis led to complete inhibition of maspin synthesis in all neutrophils.
These results suggest that estrogen and progesterone complement each other in NO-induced maspin synthesis, and do not necessarily antagonize in the synthesis of the anti-breast-cancer protein.
尽管孕激素受体（PR）状态与雌激素受体状态类似，在乳腺癌中具有预后重要性，但PR在乳腺癌中的作用仍不清楚。开展研究以确定PR在中性粒细胞中对一氧化氮诱导的maspin合成的功能，maspin是一种在非恶性乳腺细胞和循环中的中性粒细胞中产生的抗乳腺癌蛋白。
通过免疫组织化学确定PR状态。通过信使核糖核酸的体外翻译确定maspin合成，并通过酶联免疫吸附测定进行定量。通过高铁血红蛋白法测定一氧化氮。
发现中性粒细胞中的PR状态与恶性乳腺组织中的相同。孕酮与正常和PR阳性（PR+）中性粒细胞结合的Scatchard图显示，正常中性粒细胞具有11.5×10¹⁰个PR位点/细胞，解离常数（Kd）=47.619 nM，而PR+中性粒细胞具有6.6×10¹⁰个PR位点/细胞，Kd=47.619 nM。孕酮阴性（PR-）中性粒细胞未能与孕酮结合。用25 nM孕酮孵育正常和PR+中性粒细胞产生了1.317 μM一氧化氮和2.329 nM maspin；PR+中性粒细胞产生了0.72 μM一氧化氮和1.138 nM maspin。PR-中性粒细胞在孕酮存在下未能产生任何一氧化氮或maspin。抑制孕酮诱导的一氧化氮合成导致所有中性粒细胞中maspin合成完全受到抑制。
这些结果表明，雌激素和孕酮在一氧化氮诱导的maspin合成中相互补充，在抗乳腺癌蛋白的合成中不一定相互拮抗。
