华中科技大学学报（医学版）
華中科技大學學報（醫學版）
화중과기대학학보（의학판）
ACTA UNIVERSITATIS MEDICINAE TONGJI
2014年
6期
679-683
,共5页
梅花%周向东%陈贵华%蒋幼凡
梅花%週嚮東%陳貴華%蔣幼凡
매화%주향동%진귀화%장유범
丙戊酸钠%肺癌%人类M HCⅠ类分子链相关基因A%丝裂素活化蛋白激酶
丙戊痠鈉%肺癌%人類M HCⅠ類分子鏈相關基因A%絲裂素活化蛋白激酶
병무산납%폐암%인류M HCⅠ류분자련상관기인A%사렬소활화단백격매
sodium valproate%lung cancer%human M HC classⅠchain-related A%mitogen-activated protein kinase
目的：研究丙戊酸钠（sodium valproate ，VPA）作为组蛋白去乙酰酶抑制剂对肺癌A549细胞人类M HCⅠ类分子链相关基因A （human M HC classⅠchain‐related A ，M ICA ）蛋白表达的影响，并初步探讨其信号转导机制。方法使用不同浓度的VPA刺激处于对数生长期的肺癌A549细胞，培养24 h后使用RT‐PCR法检测肺癌A549细胞MICA mRNA水平，Western blot法检测MICA蛋白水平，得出最佳VPA刺激浓度。分别以丝裂素活化蛋白激酶（mitogen‐ac‐tivated protein kinase ，MAPK）通路的3条主要通路抑制剂即 ERK1／2抑制剂（PD98059）、p38蛋白激酶抑制剂（SB203580）、JNK抑制剂（SP600125）预处理 A549细胞，再将 VPA依照预定最适浓度分别加入培养24 h。使用 RT‐PCR法检测A549细胞MICA mRNA水平，Western blot法检测MICA蛋白水平。使用ELISA法检测上述各组A549细胞培养上清液中分泌型MICA（sMICA）蛋白水平。结果 RT‐PCR及Western blot检测结果显示加入VPA后A549细胞中MICA mRNA转录水平、MICA蛋白水平均显著升高（均 P＜0.05）；使用p38蛋白激酶抑制剂SB203580、ERK1／2抑制剂PD98059预处理，均能显著减弱VPA诱导的A549细胞MICA mRNA及蛋白水平的提高（P＜0.05）；各组细胞培养上清液中sMICA浓度无差异。结论 VPA能增强肺癌A549细胞MICA蛋白的表达，其机制可能与ERK、p38信号通路有关。
目的：研究丙戊痠鈉（sodium valproate ，VPA）作為組蛋白去乙酰酶抑製劑對肺癌A549細胞人類M HCⅠ類分子鏈相關基因A （human M HC classⅠchain‐related A ，M ICA ）蛋白錶達的影響，併初步探討其信號轉導機製。方法使用不同濃度的VPA刺激處于對數生長期的肺癌A549細胞，培養24 h後使用RT‐PCR法檢測肺癌A549細胞MICA mRNA水平，Western blot法檢測MICA蛋白水平，得齣最佳VPA刺激濃度。分彆以絲裂素活化蛋白激酶（mitogen‐ac‐tivated protein kinase ，MAPK）通路的3條主要通路抑製劑即 ERK1／2抑製劑（PD98059）、p38蛋白激酶抑製劑（SB203580）、JNK抑製劑（SP600125）預處理 A549細胞，再將 VPA依照預定最適濃度分彆加入培養24 h。使用 RT‐PCR法檢測A549細胞MICA mRNA水平，Western blot法檢測MICA蛋白水平。使用ELISA法檢測上述各組A549細胞培養上清液中分泌型MICA（sMICA）蛋白水平。結果 RT‐PCR及Western blot檢測結果顯示加入VPA後A549細胞中MICA mRNA轉錄水平、MICA蛋白水平均顯著升高（均 P＜0.05）；使用p38蛋白激酶抑製劑SB203580、ERK1／2抑製劑PD98059預處理，均能顯著減弱VPA誘導的A549細胞MICA mRNA及蛋白水平的提高（P＜0.05）；各組細胞培養上清液中sMICA濃度無差異。結論 VPA能增彊肺癌A549細胞MICA蛋白的錶達，其機製可能與ERK、p38信號通路有關。
목적：연구병무산납（sodium valproate ，VPA）작위조단백거을선매억제제대폐암A549세포인류M HCⅠ류분자련상관기인A （human M HC classⅠchain‐related A ，M ICA ）단백표체적영향，병초보탐토기신호전도궤제。방법사용불동농도적VPA자격처우대수생장기적폐암A549세포，배양24 h후사용RT‐PCR법검측폐암A549세포MICA mRNA수평，Western blot법검측MICA단백수평，득출최가VPA자격농도。분별이사렬소활화단백격매（mitogen‐ac‐tivated protein kinase ，MAPK）통로적3조주요통로억제제즉 ERK1／2억제제（PD98059）、p38단백격매억제제（SB203580）、JNK억제제（SP600125）예처리 A549세포，재장 VPA의조예정최괄농도분별가입배양24 h。사용 RT‐PCR법검측A549세포MICA mRNA수평，Western blot법검측MICA단백수평。사용ELISA법검측상술각조A549세포배양상청액중분비형MICA（sMICA）단백수평。결과 RT‐PCR급Western blot검측결과현시가입VPA후A549세포중MICA mRNA전록수평、MICA단백수평균현저승고（균 P＜0.05）；사용p38단백격매억제제SB203580、ERK1／2억제제PD98059예처리，균능현저감약VPA유도적A549세포MICA mRNA급단백수평적제고（P＜0.05）；각조세포배양상청액중sMICA농도무차이。결론 VPA능증강폐암A549세포MICA단백적표체，기궤제가능여ERK、p38신호통로유관。
Objective To investigate the effects of histone deacetylase inhibitor sodium valproate(VPA )on the expression of human MHC classⅠchain‐related A(MICA)protein in lung cancer A549 cells and explore its possible mechanisms of signa‐ling pathway.Methods Lung cancer A549 cells were cultured with different concentrations of VPA.The expression levels of MICA mRNA were detected by RT‐PCR and the MICA protein synthesis was judged by Western blot after cultivation for 24 h , and then optimum stimulation concentration of VPA was obtained.A549 cells were pretreated by three inhibitors of MAPK sig‐nal pathway(ERK1/2 inhibitor PD98059 ,p38 protein kinase inhibitor SB203580 ,and JNK inhibitor SP600125). Then A549 cells were stimulated with the optimum stimulation concentration of VPA.The MICA mRNA levels were analyzed by RT‐PCR ,MI‐CA protein levels assessed by Western blot ,and secreted MICA(sMICA)levels of cultural supernatants detected by ELISA after cultivation for 24 h.Results The results of RT‐PCR and Western blot showed that VPA could enhance both the mRNA and protein level of MICA in human lung cancer cell line A549(P<0.05). The ERK1/2 inhibitor PD98059 and p38 protein kinase inhibitor SB203580 could lessen the increased expression of MICA induced by VPA(P<0.05). However ,the levels of sMICA in cultural supernatants were not significantly different among the groups.Conclusion Histone deacetylase inhibitor VPA can enhance the expression of MICA in human lung cancer A549 cells via an ERK1/2 and p38 protein kinase signal related mecha‐nism.
