雌激素对肺腺癌细胞系A549中NOK表达量的影响
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摘要
目的:探讨雌激素对肺腺癌细胞系A549中NOK基因表达的影响,为研究雌激素与NOK基因在非小细胞肺癌转移中的作用提供参考依据。方法:用MTT法绘制肺腺癌细胞系A549的生长曲线,确定A549的指数增长期;用浓度梯度为1×10~(-10)-1×10~(-6)mol/L的雌激素在指数增长期刺激细胞,12 h后换液,24 h后提取总RNA及总蛋白,用q-RT-PCR及Western-blot检测各组细胞中NOK表达量。结果:A549细胞系的指数增长期为第3天;雌激素刺激细胞之后,NOK表达量普遍增高,低浓度时,NOK表达量随雌激素浓度增高而增高,高浓度时,NOK表达量随雌激素浓度增高而降低,但依然高于正常水平,组间有统计学差异,其中,当浓度为1×10~(-8)mol/L时,NOK表达量最高。结论:雌激素在一定浓度范围内可以促进肺腺癌细胞系A549中NOK基因的表达。
Objective: To study the effect of estrogen on the expression of NOK in lung adenocarcinoma cell line A549, and to provide a basis for studying the role of estrogen and NOK gene in non small cell lung cancer metastasis. Methods: MTT method was used to draw the growth curve of lung adenocarcinoma cell line A549, then determine the exponential growth phase of A549. To stimulate cells with estrogen which concentration gradient is 1×10~(-10)-1×10~(-6)mol/L in exponential growth phase. The cells were cultured in routine medium 12 h later. Then, RNA and protein were extracted. The expression of NOK was tested by q-RT-PCR and Western blot. Results:The exponential growth phase of A549 is the third day; NOK expression generally increased after estrogen stimulating, the expression of NOK increased with the increase of estrogen concentration in the low concentration, the expression of NOK decreased with the increase of estrogen concentration in the high concentration, but still higher than normal level. The NOK expression of the estrogen groups were significantly higher than that of the control group. Among them, the NOK expression was the highest when the estrogen concentration was 1×10~(-8)mol/L. Conclusion: In a certain concentration range, estrogen can promote the expression of NOK in lung adenocarcinoma cell line A549.
Objective: To study the effect of estrogen on the expression of NOK in lung adenocarcinoma cell line A549, and to provide a basis for studying the role of estrogen and NOK gene in non small cell lung cancer metastasis. Methods: MTT method was used to draw the growth curve of lung adenocarcinoma cell line A549, then determine the exponential growth phase of A549. To stimulate cells with estrogen which concentration gradient is 1×10~(-10)-1×10~(-6)mol/L in exponential growth phase. The cells were cultured in routine medium 12 h later. Then, RNA and protein were extracted. The expression of NOK was tested by q-RT-PCR and Western blot. Results:The exponential growth phase of A549 is the third day; NOK expression generally increased after estrogen stimulating, the expression of NOK increased with the increase of estrogen concentration in the low concentration, the expression of NOK decreased with the increase of estrogen concentration in the high concentration, but still higher than normal level. The NOK expression of the estrogen groups were significantly higher than that of the control group. Among them, the NOK expression was the highest when the estrogen concentration was 1×10~(-8)mol/L. Conclusion: In a certain concentration range, estrogen can promote the expression of NOK in lung adenocarcinoma cell line A549.
引文
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[14]Rivera GA,Wakelee H.Lung Cancer in Never Smokers[J].Adv Exp Med Biol,2016,893:43-57
[15]Zhou F,Jiang T,Ma W,et al.The impact of clinical characteristics on outcomes from maintenance therapy in non-small cell lung cancer:A systematic review with meta-analysis[J].Lung Cancer,2015,89(2):203-211
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[17]Cao Q,Chen M,Li Z,et al.High Novel Oncogene with Kinase-Domain(NOK)Gene Expression is Associated with the Progression of Renal Cell Carcinoma[J].Clin Lab,2016,62(1-2):179-186
[18]Orang AV,Safaralizadeh R,Hosseinpour Feizi MA,et al.Diagnostic relevance of overexpressed serine threonine tyrosine kinase/novel oncogene with kinase domain(STYK1/NOK)m RNA in colorectal cancer[J].Asian Pac J Cancer Prev,2014,15(16):6685-6689
[19]Li J,Wu F,Sheng F,et al.NOK/STYK1 interacts with GSK-3βand mediates Ser9 phosphorylation through activated Akt[J].FEBS Lett,2012,586(21):3787-3792
[20]Ding X,Jiang QB,Li R,et al.NOK/STYK1 has a strong tendency towards forming aggregates and colocalises with epidermal growth factor receptor in endosomes[J].Biochem Biophys Res Commun,2012,421(3):468-473
[2]Drake JM,Lee JK,Witte ON.Clinical targeting of mutated and wild-type protein tyrosine kinases in cancer[J].Mol Cell Biol,2014,34(10):1722-1732
[3]Liu L,Yu XZ,Li TS,et al.A novel protein tyrosine kinase NOK that shares homology with platelet-derived growth factor/fibroblast growth factor receptors induces tumorigenesis and metastasis in nude mice[J].Cancer research,2004,64(10):3491-3499
[4]Amachika T,Kobayashi D,Moriai R,et al.Diagnostic relevance of overexpressed m RNA of novel oncogene with kinase-domain(NOK)in lung cancers[J].Lung cancer,2007,56(3):337-340
[5]Moriai R,Kobayashi D,Amachika T,et al.Diagnostic relevance of overexpressed NOK m RNA in breast cancer[J].Anticancer research,2006,26(6C):4969-4973
[6]Jackson KA,Oprea G,Handy J,et al.Aberrant STYK1 expression in ovarian cancer tissues and cell lines[J].Journal of ovarian research,2009,2(1):15
[7]Chen P,Li WM,Lu Q,et al.Clinicopathologic features and prognostic implications of NOK/STYK1 protein expression in non-small cell lung cancer[J].BMC Cancer,2014,4(14):402
[8]陈鹏,张志培,金河天,等.STYK1/NOK基因转染对肺腺癌细胞迁移和侵袭的影响[J].中华肿瘤防治杂志,2011,18(8):564-567Chen Peng,Zhang Zhi-pei,Jin He-tian,et al.Impact of STYK1/NOK gene transfection on the migration and invasion of lung adenocarcinoma cell line[J].Chin J Cancer Prev Treat,2011,18(8):564-567
[9]Kimbro KS,Duschene K,Willard M,et al.A novel gene STYK1/NOK is upregulated in estrogen receptor-alpha negative estrogen receptor-beta positive breast cancer cells following estrogen treatment[J].Molecular biology reports,2008,35(1):23-27
[10]Hong QY,Wu GM,Qian GS,et al.Lung Cancer Group of Chinese Thoracic Society;Chinese Alliance Against Lung Cancer[J].Cancer,2015,121(17):3080-3088
[11]Rothschild SI.Advanced and Metastatic Lung Cancer-What is new in the Diagnosis and Therapy?[J].Praxis(Bern 1994),2015,104(14):745-750
[12]Mittal V,El Rayes T,Narula N,et al.The Microenvironment of Lung Cancer and Therapeutic Implications[J].Adv Exp Med Biol,2016,890:75-110
[13]Schwartz AG,Cote ML.Epidemiology of Lung Cancer[J].Adv Exp Med Biol,2016,893:21-41
[14]Rivera GA,Wakelee H.Lung Cancer in Never Smokers[J].Adv Exp Med Biol,2016,893:43-57
[15]Zhou F,Jiang T,Ma W,et al.The impact of clinical characteristics on outcomes from maintenance therapy in non-small cell lung cancer:A systematic review with meta-analysis[J].Lung Cancer,2015,89(2):203-211
[16]Chung S,Tamura K,Furihata M,et al.Overexpression of the potential kinase serine/threonine/tyrosine kinase 1(STYK 1)in castration-resistant prostate cancer[J].Cancer Sci,2009,100(11):2109-2114
[17]Cao Q,Chen M,Li Z,et al.High Novel Oncogene with Kinase-Domain(NOK)Gene Expression is Associated with the Progression of Renal Cell Carcinoma[J].Clin Lab,2016,62(1-2):179-186
[18]Orang AV,Safaralizadeh R,Hosseinpour Feizi MA,et al.Diagnostic relevance of overexpressed serine threonine tyrosine kinase/novel oncogene with kinase domain(STYK1/NOK)m RNA in colorectal cancer[J].Asian Pac J Cancer Prev,2014,15(16):6685-6689
[19]Li J,Wu F,Sheng F,et al.NOK/STYK1 interacts with GSK-3βand mediates Ser9 phosphorylation through activated Akt[J].FEBS Lett,2012,586(21):3787-3792
[20]Ding X,Jiang QB,Li R,et al.NOK/STYK1 has a strong tendency towards forming aggregates and colocalises with epidermal growth factor receptor in endosomes[J].Biochem Biophys Res Commun,2012,421(3):468-473