CAJ | 학술논문

간체로 보기 번체로 보기

蛋白激酶A抑制剂对HeLa细胞S期进程的影响
단백격매A억제제대HeLa세포S기진정적영향
Primary Studies on the Progression of S Phase and its Molecular Mechanism by Protein Kinase A Inhibitor in the HeLa Cell

万方数据

生物化学与生物物理进展生物化學與生物物理進展 생물화학여생물물리진전
PROGRESS IN BIOCHEMISTRY AND BIOPHYSICS
2001年 2期 232-235 ,共4页

孙霞%柳惠图%童迎凯%王端顺孫霞%柳惠圖%童迎凱%王耑順

손하%류혜도%동영개%왕단순

蛋白激酶A%蛋白激酶A抑制剂%S期%HeLa细胞蛋白激酶A%蛋白激酶A抑製劑%S期%HeLa細胞
단백격매A%단백격매A억제제%S기%HeLa세포
protein kinase A%protein kinase A inhibitor%S phase%HeLa cells

以同步化的HeLa细胞为实验材料, 研究了蛋白激酶A(PKA)抑制剂对HeLa细胞S期进程的影响及其作用的分子机理.通过TdR双阻断法, 获得了同步化的S期细胞, 3H-TdR掺入实验表明PKA抑制剂typeⅢ(80 mg/L)明显提高了S期3H-TdR的掺入水平, 提示了PKA 在S期进程中起阻抑作用.进一步实验表明，在PKA抑制剂typeⅢ作用下胸苷激酶(TK)活性和PCNA蛋白水平均有所提高，同时明显促进了CyclinA蛋白的表达, 并抑制了周期负调因子p21蛋白的水平，但对CDK2表达几乎无影响.结果表明, PKA可通过作用于PCNA和引擎分子CyclinA的水平和通过影响p21的表达负调于S期进程. 这可能是PKA负调HeLa细胞S期进程的分子机理之一 .
이동보화적HeLa세포위실험재료, 연구료단백격매A(PKA)억제제대HeLa세포S기진정적영향 급기작용적분자궤리.통과TdR쌍조단법, 획득료동보화적S기세포, 3H-TdR참입실험 표명PKA억제제typeⅢ(80 mg/L)명현제고료S기3H-TdR적참입수평, 제시료PKA 재S기 진정중기조억작용.진일보실험표명，재PKA억제제typeⅢ작용하흉감격매(TK)활성화PCNA단 백수평균유소제고，동시명현촉진료CyclinA단백적표체, 병억제료주기부조인자p21단백적 수평，단대CDK2표체궤호무영향.결과표명, PKA가통과작용우PCNA화인경분자CyclinA적수 평화통과영향p21적표체부조우S기진정. 저가능시PKA부조HeLa세포S기진정적분자궤리지일 .
The synchronized HeLa cells were used to study the effect of protein kinase A(PKA) inhibitor on the progression of S phase. Synchronized cell s in S phase were obtained by the method of TdR double block through 3H-T dR incorporation assay. The PKA inhibitor typeⅢ obviously increased the level of 3H-TdR incorporation of S phase in HeLa cells. In contrast with contro l, the activity of thymidine kinase (TK) in S phase increased, too. It indicated that PKA played an inhibitory role in S phase progression of HeLa cells. With t he method of Western blotting, the PKA inhibitor typeⅢ enhanced the level of C yclinA and PCNA, inhibited the expression of p21, which is a negative regulator of cell cycle, but had no effect on the expression of CDK2. The results showed t hat PKA could negatively regulate the S phase progression by affecting the level of CyclinA, PCNA and influencing the expression of p21 protein. This may be one of the molecular mechanisms which is involved in the negative regulation of S p hase progression by PKA in HeLa cells.