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人A549肺腺癌细胞中PTHrP的表达及与性激素状态相关性的研究(再次投稿)
Study on detecting PTHrP expression and its correlation with sex hormone status in human A549 lung adenocarcinoma cells

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DOI:
作者:
龚丽明
GONG Li-ming
作者单位:
云南省第二人民医院心胸外科
Dept.of Cardiothoracic Surgery ,The 4th Affiliated Hospital of Kunming Medical College ,The 2nd People Hospital of Yunnan Province, Kunming
关键词:
非小细胞肺癌；A549肺腺癌细胞;甲状旁腺激素相关蛋白；性激素；逆转录多聚酶链反应；适时定量PCR
non-small cell lung cancer; human A549 lung adenocarcinoma cells; parathyroid hormone–related protein (PTHrP); sex hormones; reverse transcriptase-polymerase chain reaction analysis(RT-PCR); real-time
摘要:
【摘要】 背景与目的：甲状旁腺激素相关蛋白（PTHrP）通常在非小细胞肺癌中（NSCLC）表达, 结构上与甲状旁腺激素（PTH）相似，可引起恶性肿瘤高钙血症，也能够对癌细胞产生直接作用，改变癌细胞的增殖率，调节integrin 的大小和基质金属蛋白酶的表达，故该蛋白可调节癌细胞的生长、凋亡和血管生成，这种调节作用与性激素状态相关。本研究旨在探讨人A549肺腺癌细胞中PTHrP的表达，以及雌雄激素对其表达的影响，为女性肺癌患者PTHrP的表达较男性患者PTHrP的表达高提供细胞学方面的依据，并为下一步研究PTHrP的表达与雌性肺癌裸鼠生存优势间关系而做的动物实验建模做准备。方法：人A549肺腺癌细胞培养48小时后，采用逆转录多聚酶链反应（RT-PCR）技术检测其雌激素受体（ER-α、ER-β）、雄激素受体（AR）和甲状旁腺激素相关蛋白 （PTHrP）的表达；分别用不同浓度的雌二醇和睾酮处理A549细胞48小时，用适时定量PCR（ Real-time quantitative PCR）检测其PTHrP的表达。结果：在A549细胞中雌激素受体（ER-β）、雄激素受体（AR）表达，雌激素受体（ER-α）不表达，甲状旁腺激素相关蛋白 （PTHrPmRNA）表达；用不同浓度的雌二醇及睾酮处理A549细胞48小时，雌二醇对其PTHrPmRNA的表达无影响，睾酮明显降低了PTHrPmRNA的表达。结论：人A549肺腺癌细胞中雌激素受体（ER-β）、雄激素受体（AR）表达，甲状旁腺激素相关蛋白 （PTHrP）表达；性激素可调节PTHrP的分泌，其中雄激素对PTHrP起负调节作用。
［Abstract］ Background and purpose：Parathyroid hormone–related protein (PTHrP) is commonly expressed in non–small cell lung carcinomas (non-small cell lung cancer, NSCLC). PTHrP shares structural similarities with PTH.It duplicates the effects of PTH 1-34 in tissues that bear the receptor, including causing hypercalcemia. It has a direct effect on cancer cells also that can change the proliferation rate of cancer cell, adjust the size of integrin and the expression of matrix metalloproteinase. It regulates cancer cell growth, apoptosis, and angiogenesis. This kind of effects of PTHrP is sex-dependent in lung cancer. This study examined the expression of PTHrP in human A549 lung adenocarcinoma cells and the impact of estrogen and androgen on the expression of PTHrP. It provided a cytological foundation that the expression of PTHrP in female lung cancer patients is higher than that in male patients. It prepared for further studying the relationship between the expression of PTHrP and the survival advantage of lung cancer in female nude mice. Methods：Human A549 lung adenocarcinoma cells were cultured for 48 hours. Estrogen receptor (ER-α, ER-β), androgen receptor (AR) and parathyroid hormone related protein (PTHrP) were detected using RT-PCR. Cells were treated with estradiol and testosterone with from 0.1 to 10 nM for 48 hours prior to detecting the expression of PTHrP using real-time quantitative PCR. Results：A549 cell line expressed estrogen receptor (ER-β) , androgen receptor (AR) and PTHrP based on RT-PCR analyses, but estrogen receptor (ER-α) not expressing in the cells. Cells were treated for 48 hours with from 0.1 to 10 nM estradiol, testosterone. Estradiol had no effect on the expression of PTHrPmRNA. Testosterone significantly reduced the expression of PTHrPmRNA. Conclusion：Estrogen receptor (ER-β),androgen receptor (AR) and parathyroid hormone–related protein (PTHrP) are expressed in human A549 lung adenocarcinoma cells. Sex hormones may regulate the secretion of PTHrP, in which androgen plays a negative regulatory role on PTHrP.

 

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