Estrogen regulation of TRPM8 expression in breast cancer cells.
Estrogen regulation of TRPM8 expression in breast cancer cells.
复制标题
DOI：
10.1186/1471-2407-10-212
复制
发表时间：
2010-05-19
期刊：
影响因子：
3.8
通讯作者：
Ouadid-Ahidouch H
中科院分区：
文献类型：
--
作者：
Chodon D;Guilbert A;Dhennin-Duthille I;Gautier M;Telliez MS;Sevestre H;Ouadid-Ahidouch H
钙离子通透性阳离子通道TRPM8(Melastatin相关的瞬时受体潜在成员8)在几种癌症中过度表达。本研究旨在探讨雌激素受体α(ERα)对TRPM8通道在乳腺癌中的表达、功能及其可能的调节作用。采用RT-PCR、Western印迹、免疫组织化学和siRNA技术研究TRPM8在乳腺组织中的表达，以及雌激素受体对TRPM8表达的调节作用。为了研究MCF-7细胞的通道活动，我们使用了全细胞膜片钳技术和钙离子成像技术。在乳腺癌细胞系MCF-7中，TRPM8通道在mRNA和蛋白水平上均有表达。强效TRPM8激动剂ICLIN(20μM)在去极化电位下产生强烈的外向整流电流，这与细胞内钙离子浓度升高有关，这与已建立的TRPM8通道特性一致。RT-PCR实验显示，激素剥夺48小时和72小时后，TRPM8基因的表达降低。在去掉类固醇的培养液中，加入17-β-雌二醇(E_2，10 nM)可增加TRPM8基因的表达，增加对ICLIN反应的细胞数，但不影响细胞内钙内流的幅度。此外，用小干扰α沉默ERTRPM8mRNA的表达，可降低TRPM8mRNA的表达。用免疫组织化学方法检测人乳腺组织中TRPM8通道的表达，发现乳腺癌组织中TRPM8的过度表达与肿瘤雌激素受体阳性(ER+)状态有关。综上所述，这些结果表明TRPM8通道在乳腺癌中表达并具有功能，其表达受ERα的调节。
The calcium-permeable cation channel TRPM8 (melastatin-related transient receptor potential member 8) is over-expressed in several cancers. The present study aimed at investigating the expression, function and potential regulation of TRPM8 channels by ER alpha (estrogen receptor alpha) in breast cancer. RT-PCR, Western blot, immuno-histochemical, and siRNA techniques were used to investigate TRPM8 expression, its regulation by estrogen receptors, and its expression in breast tissue. To investigate the channel activity in MCF-7 cells, we used the whole cell patch clamp and the calcium imaging techniques. TRPM8 channels are expressed at both mRNA and protein levels in the breast cancer cell line MCF-7. Bath application of the potent TRPM8 agonist Icilin (20 μM) induced a strong outwardly rectifying current at depolarizing potentials, which is associated with an elevation of cytosolic calcium concentration, consistent with established TRPM8 channel properties. RT-PCR experiments revealed a decrease in TRPM8 mRNA expression following steroid deprivation for 48 and 72 hours. In steroid deprived medium, addition of 17-beta-estradiol (E2, 10 nM) increased both TRPM8 mRNA expression and the number of cells which respond to Icilin, but failed to affect the Ca2+ entry amplitude. Moreover, silencing ERα mRNA expression with small interfering RNA reduced the expression of TRPM8. Immuno-histochemical examination of the expression of TRPM8 channels in human breast tissues revealed an over-expression of TRPM8 in breast adenocarcinomas, which is correlated with estrogen receptor positive (ER+) status of the tumours. Taken together, these results show that TRPM8 channels are expressed and functional in breast cancer and that their expression is regulated by ER alpha.
登录
查看更多内容
影响因子：
3.8
作者：
Guilbert A;Dhennin-Duthille I;Hiani YE;Haren N;Khorsi H;Sevestre H;Ahidouch A;Ouadid-Ahidouch H
通讯作者：
Ouadid-Ahidouch H
影响因子：
5.6
作者：
Borowiec, Anne-Sophie;Hague, Frederic;Ouadid-Ahidouch, Halima
通讯作者：
Ouadid-Ahidouch, Halima
影响因子：
5.5
作者：
Guilbert, Arnaud;Gautier, Mathieu;Ouadid-Ahidouch, Halima
通讯作者：
Ouadid-Ahidouch, Halima
影响因子：
4.8
作者：
Thebault, S;Lemonnier, L;Prevarskaya, N
通讯作者：
Prevarskaya, N
影响因子：
2.5
作者：
Bosch, Martha A.;Hou, Jingwen;Fang, Yuan;Kelly, Niartin J.;Ronnekleiv, Oline K.
通讯作者：
Ronnekleiv, Oline K.