Zhong Wa, Kan Fang-ju, Yu Zhong
Department of Digestive Diseases, The Second Affiliated Hospital of Sun Yat-sen University, Guangzhou.
Zhongguo Zhong Xi Yi Jie He Za Zhi. 2009 Nov;29(11):1005-8.
To study the effect of Weikangning (WKN) containing serum on growth and cell cycle regulators of gastric cancer cell.
WKN containing drug serum was prepared by gastric perfusion of WKN in different dosages to SD rats. Gastric cancer cells were cultured in medium contained the drug serum with different concentrations of WKN. The change of cell cycle was detected by flow cytometry, and the mRNA and protein expressions of CyclinD2, CyclinE, Cyclin-dependant kinase2 (CdK2), Cdk4, Cdk6 and p16(INK4a), p27(KIP1) were detected with immunohistochemistry (IHC) SABC method and RT-PCR.
After WKN intervention, the cancer cells were constrained in stage G0/G1, unable or retardatory to enter stage G (namely, the DNA synthesis stage). IHC examination showed the grey scale values of CyclinD2, CyclinE, Cdk2, Cdk4 and Cdk6 were higher, and that of p27(KIP1) and p16(INK4a) were lower in cells after moderate/high dosage WKN intervention than those in the blank control (P < 0.01); RT-PCR showed the OPTD values of CyclinD2, CyclinE, Cdk2, Cdk4 and Cdk6 were lower, and that of p27(KIP1) and p16(INK4a) were higher in cells after moderate/high dosage WKN intervention than those in the blank control (P < 0.01).
WKN can inhibit the expressions of cell cycle promoting related factors of gastric cancer cell, including CyclinD2, CyclinE, Cdk2, Cdk4 and Cdk6, also enhance the expressions of cell cycle inhibiting factors of gastric cancer cell, as p27(KIP1) and p16(INK4a), which may be the mechanism of action of the remedy in preventing the growth of gastric cancer cells.
研究胃康宁含药血清对胃癌细胞生长及细胞周期调控因子的影响。
将不同剂量胃康宁经胃灌注给SD大鼠制备含药血清。胃癌细胞培养于含不同浓度胃康宁含药血清的培养基中。采用流式细胞术检测细胞周期变化，用免疫组化（IHC）SABC法及RT-PCR检测细胞周期蛋白D2（CyclinD2）、细胞周期蛋白E（CyclinE）、细胞周期蛋白依赖性激酶2（CdK2）、细胞周期蛋白依赖性激酶4（Cdk4）、细胞周期蛋白依赖性激酶6（Cdk6）及p16（INK4a）、p27（KIP1）的mRNA和蛋白表达。
胃康宁干预后，癌细胞被阻滞于G0/G1期，不能或延迟进入S期（即DNA合成期）。免疫组化检测显示，中/高剂量胃康宁干预后细胞中CyclinD2、CyclinE、Cdk2、Cdk4和Cdk6的灰度值升高，p27（KIP1）和p16（INK4a）的灰度值降低，与空白对照组比较差异有统计学意义（P<0.01）；RT-PCR检测显示，中/高剂量胃康宁干预后细胞中CyclinD2、CyclinE、Cdk2、Cdk4和Cdk6的光密度值降低，p27（KIP1）和p16（INK4a）的光密度值升高，与空白对照组比较差异有统计学意义（P<0.01）。
胃康宁可抑制胃癌细胞周期促进相关因子CyclinD2、CyclinE、Cdk2、Cdk4和Cdk6的表达，同时增强胃癌细胞周期抑制因子p27（KIP1）和p16（INK4a）的表达，这可能是该药物抑制胃癌细胞生长的作用机制。
