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Background: The tumor cells by growth rapid, abnormal energy metabolism and self-regulation of specific proteins, the formation and survival of normal cells maintain a suitable the extracellular acidic micro-environment, and to ensure its occurrence, proliferation, invasion and metastasis. For the the acidic microenvironment generation mechanism and study its impact on the biological activity of tumor cells, may help to find out the efficiency and low toxicity means of anti-tumor, and became the new entry point for the study of anti-tumor Integrative Medicine. Mentor Professor Wei Pinkang in accordance with its long-term clinical experience gastric phlegm syndrome theory, and this theoretically founded Xiaotan Sanjie Recipe, experimental studies and clinical observations prove, can effectively inhibit gastric cancer cell growth, invasion and metastasis, to extend patient survival. Pinellia Jun Xiaotan Sanjie Recipe medicine research the ginger the Pinellia of gastric cancer cells acidic microenvironment impact, not only is the new anti-tumor mechanism to explore Xiaotan Sanjie Recipe more gastric the phlegm syndrome doctrines and tumor acidic microenvironment two 's a discussion, may help to clarify the material basis of phlegm. Purpose: starting the acidic microenvironment angle from the tumor cell survival, study the anti-tumor mechanism of the ethanol extract of ginger Pinellia and and further enrich theory of gastric sputum card. Method: 1, drugs and grouping: ginger Pinellia after pulverization, through the 95% ethanol extraction, and sterilized by filtration to obtain the ethanol extract of ginger Pinellia. Were randomly divided into 5 groups, including blank control group joined a drug-free RPMI 1640 culture medium, drug experimental group was divided into four groups, the final concentration of ethanol extract containing ginger Pinellia for 1mg/ml, 0.5mg/ml 0.25mg/ml, 0.125mg/ml. The experiment was repeated three times, the combined data for statistical. 2, SGC-7901 cells (5 × 104/m1) were seeded in three 96-well plates, each plate is divided into five groups, each group of five parallel hole, another blank zeroing hole cultured for 24, 48, 72 hours later, using the MTT method to calculate the OD values ??of the cells in each group. Calculated ginger Banxia ethanol extract 72 hours IC50. 3, each concentration of drug group intervention to the cells 72 hours after, respectively, observed under the inverted phase contrast microscope photographic. The 4 subgroups cells were cultured 0,12,24,36,48,60,72 using OD value of MTT assay of cells in each group, according to the OD value of the cell growth curve. 5 different concentrations of drug intervention cells 72 hours after AnnexinV-FITC / PI double labeling flow cytometry apoptosis of cells in each group rate. 6, use the UV spectrophotometry observed drugs after the intervention of cellular ATP enzyme activity, according to the formula (total ATP enzyme activity (U / ml) = (measuring tube OD value - control tube OD value) / (standard pipe OD value - blank tube OD value) × concentration of standard pipe (0.02umol/ml) × 6 × sample dilution factor × 7.8) calculate the concentration of drug intervention cells after 72 h of total cellular ATP enzyme activity. 7, respectively, in the drug intervention cells 0,24,48,72 hours, BCECF-AM fluorescent probe to detect cells cells within the PH value. While the extracellular medium pH value measured using a pH detector. 8, the reference drug 72 hours IC50 cells 72 hours after the intervention, the V-ATP fluorescence quantitative PCR method to detect cell gene expression of NHE1. Results: 1, different concentrations of ethanol extract of ginger Pinellia could inhibit the proliferation of human gastric cancer cell line SGC7901 cell edge burr; ginger Pinellia ethanol extract induced cell volume is reduced morphological changes, while visible cell refractive index and adherent reduced ability; Annexin V-FITC/PI double labeling assay ethanol extracts of ginger Pinellia can induce apoptosis; Total cellular ATP enzyme activity decreased significantly in the 72 hours after drug intervention; apoptosis and increased with the drug concentration rate, abnormal cell morphology changes and drugs on cell ATP enzyme inhibition showed an upward trend. 2, ginger Pinellia (0.5mg/ml) effector cells of the ethanol extract after 72 hours, the cell V - ATP, of NHE1 gene quantitative PCR inspection results are 0.174% ± 0.079%, 0.180% ± 0.165%, while the control group quantitative PCR The test results for the 0.318% ± 0.148%, 0.652% ± 0.323%. Two sets of checks by the statistics, P values ??were 0.020 and 0.001, respectively, have a statistically significant difference. 3, using BCECF-AM fluorescent probe detection can be found, ginger Pinellia ethanol extract may be provided along with the increase of drug concentration, and to intervene in the cell time extension, but also significantly reduce the the SGC7910 intracellular pH value. When intervention cell to 72 hours, outside the addition to 0.125mg/ml concentration group compared to the remaining group of drug concentration in the control group were statistically significant difference (p lt; 0.01). Extracellular pH value detection, ginger pinellia ethanol extract extracellular pH value can be increased, so that the cells inside and outside the PH value difference narrowed. Conclusions: 1, ginger Pinellia ethanol extract can inhibit the proliferation of human gastric cancer cell line SGC7901 and induce apoptosis, inhibit the enzyme activity of cellular ATP. Ethanol extract of ginger Pinellia reduce the PH value SGC7901 cells, it also increased the the PH value outside the tumor cells, reduced intracellular and extracellular pH value gradient. , Ginger pinellia ethanol extract of SGC7901 cells V-ATP, NHE1 gene expression significantly inhibited.
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