2-[哌啶乙氧基]苯基]-3-[4-羟基苯基]-2H-苯并(b)吡喃的抗肿瘤作用：涉及 GPR30/EGFR 信号通路的证据。

Anti-tumorigenic action of 2-[piperidinoethoxyphenyl]-3-[4-hydroxyphenyl]-2H-benzo(b)pyran: evidence for involvement of GPR30/EGFR signaling pathway.

机构信息

Division of Endocrinology, CSIR-Central Drug Research Institute, Lucknow-226001, UP, India.

出版信息

Gynecol Oncol. 2013 May;129(2):433-42. doi: 10.1016/j.ygyno.2013.02.005. Epub 2013 Feb 8.

DOI:10.1016/j.ygyno.2013.02.005

PMID:23402905

Abstract

OBJECTIVE

The aim of the present study was to investigate the effect of non-steroidal, pure antiestrogenic benzopyran derivative i.e., 2-[piperidinoethoxyphenyl]-3-[4-hydroxyphenyl]-2H-benzo(b)pyran (K-1) on the growth of human endometrial cancer cells in vivo and in vitro and to elucidate its mechanism of action.

METHODS

Cell proliferation was assayed by measuring the incorporation of 5'-bromo-2'-deoxyuridine in Ishikawa and primary endometrial cancer cells. The expression of proliferation and apoptotic markers was analyzed by immunoblotting. The effect of K-1 on GPR30-regulated proteins was analyzed by ELISA and by immunoblotting. Nude mice bearing subcutaneous implanted-Ishikawa tumors, were treated for 14days with K-1 (200μg/kg body weight/day/orally). The proliferation markers, GPR30-regulated proteins and apoptotic markers were analyzed by immunoblotting in tumor xenograft. The apoptotic effect of compound K-1 was determined by TUNEL assay.

RESULTS

Compound K-1 inhibited proliferation of endometrial adenocarcinoma cells and decreased the expression of proliferation markers. It caused apoptosis by increasing the expression of apoptotic markers (NOXA, PUMAα) and reducing the expression of p-CREB and BclxL. Compound interfered with GPR30-regulated-EGFR activation, decreased p-ERK, p-c-jun, c-fos, cyclinD1 and c-myc expression. Treatment of tumor-bearing mice with K-1 resulted in a significant decrease in tumor volume and weight. Decreased expression of p-ERK and its downstream molecules and increased expression of apoptotic markers were observed in tumor in K-1 treated animals.

CONCLUSION

Findings suggest the potent inhibitory effect of compound K-1 on endometrial cancer cellular growth (in-vitro) and on tumor size (in-vivo) which is mediated at least, in part, by interference with GPR30-signaling.

摘要

目的

本研究旨在探讨非甾体、纯抗雌激素苯并吡喃衍生物 2-[哌啶乙氧基苯]-3-[4-羟基苯基]-2H-苯并（b）吡喃（K-1）对体内和体外人子宫内膜癌细胞生长的影响，并阐明其作用机制。

方法

通过测量 5'-溴-2'-脱氧尿苷在 Ishikawa 和原发性子宫内膜癌细胞中的掺入来测定细胞增殖。通过免疫印迹分析增殖和凋亡标志物的表达。通过 ELISA 和免疫印迹分析 K-1 对 GPR30 调节蛋白的影响。用 K-1（200μg/kg 体重/天/口服）治疗携带皮下植入 Ishikawa 肿瘤的裸鼠 14 天。通过免疫印迹分析肿瘤异种移植物中的增殖标志物、GPR30 调节蛋白和凋亡标志物。通过 TUNEL 测定法确定化合物 K-1 的凋亡作用。

结果

化合物 K-1 抑制子宫内膜腺癌细胞的增殖并降低增殖标志物的表达。它通过增加凋亡标志物（NOXA、PUMAα）的表达和降低 p-CREB 和 BclxL 的表达来引起凋亡。化合物干扰 GPR30 调节的 EGFR 激活，降低 p-ERK、p-c-jun、c-fos、cyclinD1 和 c-myc 的表达。用 K-1 治疗荷瘤小鼠导致肿瘤体积和重量显著减小。在 K-1 处理的动物中观察到肿瘤中 p-ERK 及其下游分子的表达减少和凋亡标志物的表达增加。