1.Effect of weikangning on growth and cell cycle regulators of gastric cancer cell.
Wa ZHONG ; Fang-ju KAN ; Zhong YU
Chinese Journal of Integrated Traditional and Western Medicine 2009;29(11):1005-1008
OBJECTIVETo study the effect of Weikangning (WKN) containing serum on growth and cell cycle regulators of gastric cancer cell.
METHODSWKN 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.
RESULTSAfter 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).
CONCLUSIONWKN 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.
Animals ; Cell Cycle ; drug effects ; Cell Cycle Proteins ; metabolism ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Drugs, Chinese Herbal ; pharmacology ; Gene Expression Regulation, Neoplastic ; Male ; Rats ; Rats, Sprague-Dawley ; Serum ; chemistry ; Stomach Neoplasms ; metabolism ; pathology
2.Effects of weikangning contained serum on growth of gastric cancer cell, expression of vascular endothelial growth factor and its receptors including KDR and fit-1.
Qing-ming LI ; Fang-ju KAN ; Cun-yun MIN
Chinese Journal of Integrated Traditional and Western Medicine 2006;26 Suppl():88-92
OBJECTIVETo observe the effect of drug-containing serum of Chinese traditional herbal decoction Weikangning (WKN) on growth of gastric cancer cell, expression of vascular endothelial growth factor (VEGF) and its receptors including KDR and Flt-1.
METHODSA total of 120 male Wistar rats were given high, medium and low-dose WKN. After the drug-containing serum was prepared, the gastric cancer cells MGC-803 of different dose groups were cultured with the drug-containing serum, respectively. The gastric cell growth was observed by using light microscope and flow cytometer,the expression of VEGF and its receptor Flt-1 was detected with SABC immunohistochemistry method and the mRNA expression levels of VEGF and its receptors including KDR and Flt-1 of different groups were detected with reverse transcription-polymerase chain reaction (RT-PCR), respectively.
RESULTSThe gastric cancer cell growth and cell cycle of the three medicated groups were significantly improved as compared with those of the control group (P <0. 01) ,the proportion of cells in G0 - G1 phase was increased,while the cells in S phase were decreased. It was shown that the apoptotic rates were increased in the medicated groups in a dose-dependent manner. The gray scales ( microm(2) ) of VEGF and Flt-1 in high, medium and low dose groups was 182. 44 +/-0. 54,178. 65 +/-0. 56,174. 80 +/-0. 81 and 168. 51 +/- 0. 81,162. 01 +/-0. 52,148. 20 +/-0. 69, respectively vs 147.82 +/-0. 15(P <0.01) and 144.31 +/-0.71 (P <0.01) in the control group. The mRNA expression of VEGF and its receptors significantly decreased in gastric cancer cells after cultured with WKN contained serum (P < 0. 01 ).
CONCLUSIONWKN has inhibitory effects on gastric cancer cell growth and mRNA expression of VEGF as well as its receptors KDR and Flt-1.
Animals ; Antineoplastic Agents, Phytogenic ; pharmacology ; therapeutic use ; Cell Cycle ; drug effects ; Cell Proliferation ; drug effects ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Male ; Rats ; Rats, Wistar ; Stomach Neoplasms ; drug therapy ; enzymology ; metabolism ; Vascular Endothelial Growth Factor A ; metabolism ; Vascular Endothelial Growth Factor Receptor-1 ; metabolism ; Vascular Endothelial Growth Factor Receptor-2 ; metabolism