Guilbert Arnaud, Dhennin-Duthille Isabelle, Hiani Yassine El, Haren Nathalie, Khorsi Hafida, Sevestre Henri, Ahidouch Ahmed, Ouadid-Ahidouch Halima
Laboratoire de Physiologie Cellulaire et Moléculaire, JE Canaux ioniques dans le cancer du sein, Faculté des Sciences, Université de Picardie Jules Verne, 33 Rue St Leu 80039, Amiens, France.
BMC Cancer. 2008 May 2;8:125. doi: 10.1186/1471-2407-8-125.
TRP channels have been shown to be involved in tumour generation and malignant growth. However, the expression of these channels in breast cancer remains unclear. Here we studied the expression and function of endogenous TRPC6 channels in a breast cancer cell line (MCF-7), a human breast cancer epithelial primary culture (hBCE) and in normal and tumour breast tissues.
Molecular (Western blot and RT-PCR), and immunohistochemical techniques were used to investigate TRPC6 expression. To investigate the channel activity in both MCF-7 cells and hBCE we used electrophysiological technique (whole cell patch clamp configuration).
A non selective cationic current was activated by the oleoyl-2-acetyl-sn-glycerol (OAG) in both hBCE and MCF-7 cells. OAG-inward current was inhibited by 2-APB, SK&F 96365 and La3+. TRPC6, but not TRPM7, was expressed both in hBCE and in MCF-7 cells. TRPC3 was only expressed in hBCE. Clinically, TRPC6 mRNA and protein were elevated in breast carcinoma specimens in comparison to normal breast tissue. Furthermore, we found that the overexpression of TRPC6 protein levels were not correlated with tumour grades, estrogen receptor expression or lymph node positive tumours.
Our results indicate that TRPC6 channels are strongly expressed and functional in breast cancer epithelial cells. Moreover, the overexpression of these channels appears without any correlation with tumour grade, ER expression and lymph node metastasis. Our findings support the idea that TRPC6 may have a role in breast carcinogenesis.
瞬时受体电位(TRP)通道已被证明与肿瘤发生和恶性生长有关。然而,这些通道在乳腺癌中的表达仍不清楚。在此,我们研究了内源性TRPC6通道在乳腺癌细胞系(MCF-7)、人乳腺上皮原代培养物(hBCE)以及正常和肿瘤乳腺组织中的表达及功能。
采用分子生物学技术(蛋白质免疫印迹法和逆转录聚合酶链反应)以及免疫组织化学技术研究TRPC6的表达。为研究MCF-7细胞和hBCE中的通道活性,我们采用了电生理技术(全细胞膜片钳模式)。
油酰-2-乙酰基-sn-甘油(OAG)在hBCE和MCF-7细胞中均可激活非选择性阳离子电流。OAG内向电流可被2-氨基乙氧基二苯硼酸(2-APB)、SK&F 96365和La3+抑制。TRPC6而非TRPM7在hBCE和MCF-7细胞中均有表达。TRPC3仅在hBCE中表达。临床上,与正常乳腺组织相比,乳腺癌标本中TRPC6的信使核糖核酸(mRNA)和蛋白质水平升高。此外,我们发现TRPC6蛋白水平的过表达与肿瘤分级、雌激素受体表达或淋巴结阳性肿瘤无关。
我们的结果表明,TRPC6通道在乳腺癌上皮细胞中强烈表达且具有功能。此外,这些通道的过表达似乎与肿瘤分级、雌激素受体表达和淋巴结转移均无关联。我们的研究结果支持TRPC6可能在乳腺癌发生过程中起作用这一观点。