中国病理生理杂志
中國病理生理雜誌
중국병리생리잡지
CHINESE JOURNAL OF PATHOPHYSIOLOGY
2015年
8期
1451-1456
,共6页
肺腺癌%DNA甲基化%细胞凋亡%地西他滨
肺腺癌%DNA甲基化%細胞凋亡%地西他濱
폐선암%DNA갑기화%세포조망%지서타빈
Lungadenocarcinoma%DNAmethylation%Apoptosis%Decitabine
目的:研究肺腺癌细胞P15中凋亡相关基因甲基化状态与化疗敏感性的关系。方法:采用甲基化特异性PCR法检测未处理对照组和地西他滨( DAC)处理组P15细胞的 p73、p14ARF、p16INK4a和bax基因甲基化情况,利用RT-PCR检测p73、bcl-xL、bad、bax、p14ARF和p16INK4a的mRNA表达;采用平板克隆形成实验和细胞生长抑制实验检测DAC处理前后P15细胞对顺铂( C-DDP)的敏感性,DAPI染色法检测DAC处理前后C-DDP对P15细胞的凋亡情况。结果:p73、p16INK4a和bax在甲基化状态下均有表达,经DAC处理后p16INK4a表达减弱,p73和bax表达消失。 p73、p16INK4a和bax在非甲基化状态下表达较弱,经DAC处理后三者均表达增强。 DAC加C-DDP处理组与未处理对照组相比,前者的克隆形成率和细胞生存显著下降( P<0.05)。 DAC加C-DDP组的细胞凋亡显著高于未处理对照组( P<0.05)。结论:由于DAC活化多个因甲基化而表达沉默的促凋亡基因,用DAC处理肺腺癌细胞P15后,P15细胞对化疗药物C-DDP的敏感性增强。 DAC和C-DDP协同作用促进了肿瘤细胞的凋亡。 DAC与C-DDP联合应用可逆转化疗耐药,恢复肺腺癌细胞对化疗药物的敏感性,两者具有明显的协同抗肿瘤作用。
目的:研究肺腺癌細胞P15中凋亡相關基因甲基化狀態與化療敏感性的關繫。方法:採用甲基化特異性PCR法檢測未處理對照組和地西他濱( DAC)處理組P15細胞的 p73、p14ARF、p16INK4a和bax基因甲基化情況,利用RT-PCR檢測p73、bcl-xL、bad、bax、p14ARF和p16INK4a的mRNA錶達;採用平闆剋隆形成實驗和細胞生長抑製實驗檢測DAC處理前後P15細胞對順鉑( C-DDP)的敏感性,DAPI染色法檢測DAC處理前後C-DDP對P15細胞的凋亡情況。結果:p73、p16INK4a和bax在甲基化狀態下均有錶達,經DAC處理後p16INK4a錶達減弱,p73和bax錶達消失。 p73、p16INK4a和bax在非甲基化狀態下錶達較弱,經DAC處理後三者均錶達增彊。 DAC加C-DDP處理組與未處理對照組相比,前者的剋隆形成率和細胞生存顯著下降( P<0.05)。 DAC加C-DDP組的細胞凋亡顯著高于未處理對照組( P<0.05)。結論:由于DAC活化多箇因甲基化而錶達沉默的促凋亡基因,用DAC處理肺腺癌細胞P15後,P15細胞對化療藥物C-DDP的敏感性增彊。 DAC和C-DDP協同作用促進瞭腫瘤細胞的凋亡。 DAC與C-DDP聯閤應用可逆轉化療耐藥,恢複肺腺癌細胞對化療藥物的敏感性,兩者具有明顯的協同抗腫瘤作用。
목적:연구폐선암세포P15중조망상관기인갑기화상태여화료민감성적관계。방법:채용갑기화특이성PCR법검측미처리대조조화지서타빈( DAC)처리조P15세포적 p73、p14ARF、p16INK4a화bax기인갑기화정황,이용RT-PCR검측p73、bcl-xL、bad、bax、p14ARF화p16INK4a적mRNA표체;채용평판극륭형성실험화세포생장억제실험검측DAC처리전후P15세포대순박( C-DDP)적민감성,DAPI염색법검측DAC처리전후C-DDP대P15세포적조망정황。결과:p73、p16INK4a화bax재갑기화상태하균유표체,경DAC처리후p16INK4a표체감약,p73화bax표체소실。 p73、p16INK4a화bax재비갑기화상태하표체교약,경DAC처리후삼자균표체증강。 DAC가C-DDP처리조여미처리대조조상비,전자적극륭형성솔화세포생존현저하강( P<0.05)。 DAC가C-DDP조적세포조망현저고우미처리대조조( P<0.05)。결론:유우DAC활화다개인갑기화이표체침묵적촉조망기인,용DAC처리폐선암세포P15후,P15세포대화료약물C-DDP적민감성증강。 DAC화C-DDP협동작용촉진료종류세포적조망。 DAC여C-DDP연합응용가역전화료내약,회복폐선암세포대화료약물적민감성,량자구유명현적협동항종류작용。
AIM:Toinvestigatetheassociationbetweenmethylationstatusofapoptosis-relatedgenesandche-mosensitivity in the lung adenocarcinoma cell line P 15.METHODS: Methylation-specific PCR was applied to detect the methylation status of p73, p14ARF, p16INK4a and bax genes of P15 cells in untreated control group and decitabine (DAC) treatment group.RT-PCR was used to detect the expression of p 73, bcl-xL, bad, bax, p14ARF and p16INK4a at mRNA level. Colony formation assay and cell growth inhibition assay were used to detect the sensitivity of P 15 cells to cis-diaminedichlo-roplatinum ( C-DDP) before and after DAC treatment .DAPI staining was used to determine the apoptosis of P 15 cells ex-posed to C-DDP before and after DAC treatment .RESULTS:p73, p16INK4a and bax were expressed in the methylation sta-tus.After DAC treatment, p16INK4a expression was decreased , and the expression of p73 and bax disappeared .The expres-sion of p73, p16INK4a and bax in the unmethylated status was weak , but the enhanced expression was observed following DAC treatment.After P15 cells were treated with DAC and C-DDP, the colony formation rate of the P15 cells was signifi-cantly decreased as compared with untreated control group .The apoptotic P15 cells in DAC+C-DDP treatment group were significantly higher than those in untreated control group (P<0.05).CONCLUSION:After treated with DAC, the sensi-tivity of P15 cells to C-DDP is increased due to the activation of silenced pro-apoptotic genes .DAC and C-DDP synergisti-cally promote tumor cell apoptosis .They have significant anti-tumor effect .