The Expression Of Various Progesterone Relevant Receptors In Four Lung Adenocarcinoma Cell Lines

Objective:In recent years, the relationship between the occurrence and development of non-small cell lung cancer (NSCLC) and sex hormone have been attracting the researchers attention. Estrogen can promote the development of lung cancer, whereas, mediated by different receptors, progesterone produce positive or negative effects on lung cancer. In this study, we selected the lung adenocarcinoma A549cell line, A549/DDP (cisplatin-resistant A549cell line), lung adenocarcinoma PC-9cell line and PC-9/GR (gefitinib-resistant PC-9cell line) to detect the expression of various progesterone relevant receptors (mPRα、mPRβ、 mPRγ、PR-A/B、PR-B、PGRMC1and PGRMC2) and to further analysis the relationship between mPRa expression and drug resistance.Methods:①On the mRNA level, Used real-time PCR technology to detect the expression of seven kinds of relevant progesterone receptor (mPRa, mPRp, mPRy, PR-A/B, PR-B, PGRMC1and PGRMC2) in the four kinds of lung adenocarcinoma cell lines (A549, A549/DDP, PC-9, PC-9/GR);②Western Blot techniques quantified the expression of mPRa protein in the four different lung adenocarcinoma cell lines;③ Cell immunofluorescence technique to detect the location of mPRa in the four kinds of lung adenocarcinoma cell line.Results:(1) At the mRNA level, the same kind of progesterone receptors in four different lung adenocarcinoma cell lines expressed obvious differences(P<0.05):mPRa and PGRMC2were the highest expression in PC-9cell line. mPRβ、mPRγ、PR-A/B、PR-B and PGRMC1were the highest expression in A549/DDP cell line. On the other hand, mPRa was the lowest expression in A549/DDP cell line. mPRβ、PR-A/B and PGRMC2were the lowest expression in A549cell line. mPRγ、PR-B and PGRMC1were the lowest expression in PC-9/GR cell line.(2) Also at the mRNA level, the same kind of lung adenocarcinoma cell lines expressed with significant differences on the seven different progesterone receptors(P<0.01):Both the A549and A549/DDP cell lines were found mPRy expressed highest and PR-A/B expressed lowest. Both PC-9and PC-9/GR cell lines were found PGRMC2expressed highest and PR-A/B expressed lowest.(3) Both in mRNA level detected by FQ-PCR and protein level detected by Western Blot were found that, compared with their parental cell lines (A549、PC-9), drug-resistant lung adenocarcinoma cell lines (A549/DDP、PC-9/GR) were down-regulated their expression of mPRa.(4) In morphology, cell immunofluorescence showed that all four kinds of lung adenocarcinoma cancer cell lines had clearly visible mPRa expression. As in high-power microscope, it showed that mPRa was expressed in the cell membrane.Conclusions:①All seven kinds of relevant progesterone receptor (mPRα, mPRβ, mPRγ, PR-A/B, PR-B, PGRMC1and PGRMC2) were expressed in the four kinds of lung adenocarcinoma cell lines (A549, A549/DDP, PC, PC-9of PC-9/GR);②At the mRNA level, the same kind of progesterone receptors in four different lung adenocarcinoma cell lines expressed obvious differences and the same kind of lung adenocarcinoma cell lines expressed with significant differences on each of the seven different progesterone receptors;③The expression of mPRa in lung adenocarcinoma cell lines associated with drug resistance;④The expression of mPRa in the four kinds of lung adenocarcinoma cancer cell line was located in the cell membrane.