[Effect of weikangning on growth and cell cycle regulators of gastric cancer cell].

OBJECTIVE: To study the effect of Weikangning (WKN) containing serum on growth and cell cycle regulators of gastric cancer cell. METHODS: 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. RESULTS: 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). CONCLUSION: 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.

[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].

OBJECTIVE: To 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. METHODS: A 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. RESULTS: The 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 ). CONCLUSION: WKN has inhibitory effects on gastric cancer cell growth and mRNA expression of VEGF as well as its receptors KDR and Flt-1.

[Effect of weikangning decoction on expression of cyclin E, CDK2, and p27].

OBJECTIVE: To observe the effect of containing drugs serum of weikanguing decoction (WKN) on expression of Cyclin E, CDK2 (Cyclin dependant kinase 2, CDK2) and p27 in gastric cancer cell line MGC-803. METHODS: Total of 120 male Wistar rats were divided into control group, high dose group, medium dose group and low dose group fed with natural saline, 20,10, and 5 g/kg of WKN, respectively. The experimental animals were finally killed for the preparation of drug-containing serum. The gastric cancer cell MGC-803 was cultured with the drug-containing serum drawn from the rats in different groups. The expression of Cyclin E, CDK2 and p27 was detected with immunohistochemistry method-SABC. The expression of mRNA of Cyclin E, CDK2 and p27 were detected with RT-PCR. RESULTS: In high, medium and low dose groups, the gray scales of cyclin E, CDK2 and the OPDTI values of p27 increased significantly, but the gray scales of p27 and the OPDTI values of cyclin E, CDK2 decreased remarkble, compared with that in control group. CONCLUSION: The decoction of WKN can decrease the expression of Cyclin E, CDK2, increases the expression of p27. This effect may be involved in mechanism of gastric cancer cell growth inhibition induced by WKN.

[Effects of Naohuandan Recipe on learning and memory abilities of SAM-P/8 mice and its role in anti-oxidation and anti-apoptosis].

OBJECTIVE: To investigate the effects of Naohuandan Recipe on learning and memory abilities of SAM-P/8 mice and its role in anti-oxidation and anti-apoptosis. METHODS: Forty SAM-P/8 mice were randomly divided into four groups, which were untreated (normal saline-treated) group, Yinkeluo Tablets (extracts of gingko leaf)-treated group, low-dose Naohuandan Recipe-treated group and high-dose Naohuandan Recipe-treated group. Mice in these groups were given corresponding drugs orally for 105 days. Then the performances of learning and memory of mice were tested by a step-down passive avoidance task and a Y-maze test. The serum levels of superoxide dismutase (SOD), malondialdehyde (MDA) and glutathione peroxidase (GSH-Px) were detected. The expression level of bcl-xl mRNA in cerebral cortex and hippocampus of mice was detected by reverse transcription-polymerase chain reaction (RT-PCR). RESULTS: The performances of learning and memory in the Yinkeluo Tablets-treated group, low- and high-dose Naohuandan Recipe-treated groups were significantly improved as compared with those in the untreated group (P<0.05 or P<0.01), and such performance was the best in the high-dose Naohuandan Recipe-treated group among these four groups (P<0.01). The serum levels of SOD and GSH-Px and the expression of bcl-xl mRNA in cerebral cortex and hippocampus of mice in the Yinkeluo Tablets-treated group, low- and high-dose Naohuandan Recipe-treated groups were also significantly higher than those in the untreated group (P<0.05 or P<0.01), while the serum level of MDA in the untreated group was higher than that in the other three groups (P<0.01). CONCLUSION: Naohuandan Recipe can improve learning and memory abilities of SAM-P/8 mice, and this effect may be related to its anti-oxidation efficacy and enhancement of expression level of bcl-xl mRNA.

Effect of Weikangning on gastric cancer cell growth and expression of vascular endothelial growth factor and its receptors KDR and Flt-1.

AIM: To observe the effect of Chinese traditional herbal decoction Weikang-ning (WKN) on cell growth and expression of VEGF and its receptors KDR and Flt-1 in gastric cancer cell line MGC-803. METHODS: A total of 120 male Wistar rats were divided into control group, high dose, medium dose and low dose groups fed with natural saline, 20, 10, and 5 g/kg of WKN, respectively. The experimental animals were finally killed for the preparation of drug-containing serum. The gastric cancer cell MGC-803 was cultured with the drug-containing serum drawn from the rats in different groups. We observed the growth condition of the cancer cells with light microscope and flow cytometer. The expression of mRNA of VEGF and its receptors KDR and Flt-1 was detected with RT-PCR. RESULTS: The proportion of cells in G(0)-G(1) phase was (65.40+/-0.41)%, (56.92+/-0.62)%, (55.89+/-0.69)% in high, medium and low dose groups respectively vs (41.35+/-0.55)% in control group (P<0.01), while the cells in G(2)-S and S phases were (11.62+/-0.62)% and (22.99+/-0.69)%, (17.08+/-0.80)% and (26.00+/-0.71)%, (19.37+/-0.57)% and (24.74+/-0.64)% in high, medium and low dose groups, respectively, vs (23.65+/-0.56)% and (35.00+/-0.60)% in control group (P<0.01). The expression of mRNA of VEGF and its receptors was significantly decreased, the area of electrophoresis bands (AREA), the absorptivity of mean optical density (A) and the product of AREA and A were significantly lower in WKN-administered groups than that in control group (P<0.01). CONCLUSION: The decoction of WKN suppresses the growth of gastric cancer cell MGC-803 and decreases the expression of mRNA of both VEGF and its receptors KDR and Flt-1.