CAJ | 학술논문

目的观察蛋白酪氨酸激酶Pyk2在胃癌组织中的表达,探讨Pyk2在胃癌中的作用机制.方法应用Western blot和荧光定量PCR方法检测正常胃黏膜组织和胃癌组织中Pyk2表达的差异.应用质粒转染的方法,将质粒PKH3-Pyk2和PKH3-PKM(PKM,dominant-negative Pyk2 mutant)转染胃癌细胞系BGC-803,观察凋亡相关蛋白bcl-2和bax的变化,探讨Pyk2在胃癌中作用机制.结果 Western blot结果显示Pyk2蛋白在28例正常胃黏膜组织中均有表达,在胃癌组织中表达水平很低或几乎无表达,差异有统计学意义(t=3.53,P＜0.05).荧光定量PCR结果显示:胃癌组织中Pyk2 mRNA表达水平明显低于正常胃黏膜组织,差异有统计学意义(t=8.02,P＜0.05).转染PKH3-Pyrk2质粒后,对凋亡抑制蛋白bcl-2影响不明显,而促凋亡蛋白bax的表达升高.结论 Pyk2在正常胃黏膜组织中高表达,在胃癌组织中低表达.Pylc2可能通过上调bax蛋白的表达而促进胃癌细胞凋亡.
목적 관찰단백락안산격매Pyk2재위암조직중적표체,탐토Pyk2재위암중적작용궤제.방법 응용Western blot화형광정량PCR방법검측정상위점막조직화위암조직중Pyk2표체적차이.응용질립전염적방법,장질립PKH3-Pyk2화PKH3-PKM(PKM,dominant-negative Pyk2 mutant)전염위암세포계BGC-803,관찰조망상관단백bcl-2화bax적변화,탐토Pyk2재위암중작용궤제.결과 Western blot결과현시Pyk2단백재28례정상위점막조직중균유표체,재위암조직중표체수평흔저혹궤호무표체,차이유통계학의의(t=3.53,P＜0.05).형광정량PCR결과현시:위암조직중Pyk2 mRNA표체수평명현저우정상위점막조직,차이유통계학의의(t=8.02,P＜0.05).전염PKH3-Pyrk2질립후,대조망억제단백bcl-2영향불명현,이촉조망단백bax적표체승고.결론 Pyk2재정상위점막조직중고표체,재위암조직중저표체.Pylc2가능통과상조bax단백적표체이촉진위암세포조망.
Objective To investigate whether Pyk2 is expressed differently in normal gastric mucosas and gastric carcinoma tissues.then study the mechanism of Pyk2 in gastric carcinoma cell lines preliminarily.Methods Western blot assay and fluorescent quantitation PCR were used to detect the expression of Pyk2 in 28 normal gastric mucosas and gastric carcinoma tissues.Plasmids contalning both wild types Pyk2 full length as well as that bearing PKM(dominant-negative Pyk2 mutant)were transfected into gastric carcinoma cell lines to observe the influence of Pvk2 on protein related apoptosis:bcl-2 and bax.Resuits Western blot assay showed that the expression of Pvk2 protein was high in normal gastric mucosas,while was low or almost disappeared in gastric carcinoma tissues.The difierence was significant(t=3.53.P＜0.05).Fluorescent quantitation PCR also showed that the expression of Pyk2 mRNA in gastric carcinoma tissues was lower than that in normal gastric mucosas with the difference being significant (t=8.02,P＜0.05).After transfection of different plasmids into BGC-803 gastric carcinoma cells,no obvious changes of anti-apoptotic protein bcl-2 were observed.while the level of bax protein increased.Condusion Pyk2 protein is expressed highly in normal gastric mucosas,whereas its expression is declined significantly in gastric carcinoma tissues.Pyk2 may induce apoptosis of gastric carcinoma cell through up-resulating the expression of bax protein.