| Background and purposeThe prevention and treatment of malignant tumor have become difficulty and key the scholars face as a result of the high incidence and mortality of cancer. The treatment of malignant tumor has experienced a long process. Surgical resection from extensive to the organization and function of the conservative therapy, have been developed to the current preventive surgery, minimally invasive treatment and individual therapy. Medical treatment has been developed from the nonspecific cytotoxic drugs therapy, the different stages of the cancer cells of high-dose drug killer treatment, to targeting therapy, biotherapy, etc. Accompanied with other therapies, radiotherapy can be less likely to damage the healthy tissue and improve the curative effect. When the tumor was found, most of them were already the middle-late stage. The middle-late tumor patients who were medically inoperable were generally offered chemical treatment. The patients of the long-term chemotherapy were found to have drug resistance, more side effects, higher cost and lower the quality of life. The development and application of the nature medicines has become the research focus these years. The anti-tumor mechanism of traditional Chinese medicine may inhibit the growth of tumor cells and induce apoptosis through affecting the different phase of tumor cell generation cycle, which is the viewpoint of strengthening body resistance and eliminating evil. Compared with western medicine, traditional Chinese medicine has extensive sources, low price and slight side effects. Traditional Chinese medicine can be researched through single drug or compound mixture, withdrawing its anti-tumor components, accessing to new anticancer drugs, and opening new avenues for the treatment of tumors.Rhizoma polygonati, Morinda officinalis, radix angelicae dahuricae are the traditional Chinese medicine, being extracted the main component for the anti-tumor research. This experiment chooses human esophageal carcinoma Eca-109cell, human lung cancer HGC-27cell, human colon cancer HCT-8cell as experimental objects.In this study, deal with three types of cells with different concentrations and different combinations of Chinese herb extracts in vitro, then detect cell biological activity, changes in cell morphology and cell cycle. These experiments will provide further theoretical basis for the experimental study of follow-up entity metastases.Methods1Drug preparation According to the literature, the drug in the extraction method to extract anti-tumor components of crude extracting from all kinds of medicine。 Three kinds of Chinese traditional medicine antitumor composition mainly for Rhizoma polygonati polysaccharide, morinda officinalis how liquid, radix angelicae dahuricae isoimperatorin. Major composition dissolved in the experiment for different drug concentration. Drug concentration=Traditional Chinese medicine weight/medium volume.2Cell culture Eca-109, HGC-27, HCT-8cells used to contain10%inactivated fetal bovine serum RPMI-1640medium, at37℃and routinely cultured in5%CO2incubator.24to48hour for liquid,48-72hour passage.3The determination of the inhibition rates Take the Eca-109, HGC-27, HCT-8cells in the logarithmic growth phase,5x104/ml concentration of cells inoculated in96-well plates, each well inoculated with200μL, cultured36h, set in the control group (no dosing)and experimental groups, each group of six parallel hole, at37℃ and routinely cultured in5%CO2incubator, after48h, thiazolyl blue (MTT) reduction method to determine the growth inhibitory rate of cells of each experimental group. The experiment was repeated three times (n=18).4Watch cell morphology After drug action in Eca-109cells48hours, observed the morphological changes using an inverted microscope.5Flow cytometry to detect cell cycle6Statistical analysis statistical software SPSS17.0analysis, single factor analysis of variance test with a=0.05for the inspection standards.Results1Inhibitory effect of Rhizoma polygonati,morinda officinalis how,radix angelicae dahuricae on the three tumor cells.1.1Eca-109cell growth of inhibition when Rhizoma polygonati concentration was20mg/ml, the inhibition rate reached56.69%, when Morinda officinalis how concentration was20mg/ml, the inhibition rate reached61.66%,when radix angelicae dahuricae concentration was4mg/ml, the inhibition rate of24.27%,when three drugs combined, the highest inhibition rate of88.31%.1.2HGC-27cell growth of inhibition when Rhizoma polygonati concentration was20mg/ml, the inhibition rate reached51.65%, when Morinda officinalis how concentration was20mg/ml, the inhibition rate reached56.96%,when radix angelicae dahuricae concentration was4mg/ml, the inhibition rate of23.84%,when three drugs combined, the highest inhibition rate of87.96%.1.3HCT-8cell growth of inhibition when Rhizoma polygonati concentration was20mg/ml, the inhibition rate reached58.19%, when Morinda officinalis how concentration was20mg/ml, the inhibition rate reached60.19%,when radix angelicae dahuricae concentration was4mg/ml, the inhibition rate of28.27%,when three drugs combined, the highest inhibition rate of87.70%.2Blank control group:Eca-109cell morphology have a polygon, and the adherent growth, growth in good condition. Rhizoma polygonati (lmg/ml) and radix angelicae dahuricae (0.5mg/ml):Most of the adherent cells lose their normal morphology, cell shape into a round and less refraction. Triple set group (the Rhizoma polygonati4mg/ml+the Morinda officinalis how4mg/ml+the radix angelicae dahuricae0.5mg/ml):into a group of cells separated from each other, cohesion and narrow cells, cell membrane becomes wrinkled, nuclear chromatin deeply stained pyknotic gathered in the nuclear the edge of the membrane, may be more common in the vacuole deformation.3、Eca-109cells, HGC-27cells and HCT-8cells were cultured in vitro,by different single-agent and combination therapy, the percentage rate of cells in S phase of the experimental group significantly increased,and in G0/G1phase cells have the percentage rate significant decrease; cells were blocked in S phase and inhibition of apoptosis.Conclusions1Rhizoma polygonati, Morinda officinalis, radix angelicae dahuricae three kinds of traditional Chinese medicine can significantly inhibit Eca-109cells, HGC-27cells and HCT-8cell growth activity. And a combination of drugs used in conjunction, a more pronounced inhibition of cell growth activity.2Rhizoma polygonati, Morinda officinalis, radix angelicae dahuricae three kinds of traditional Chinese medicine let Eca-109cells, the HGC-27cells and HCT-8cells arrest in S phase and the role of three-drug combination can induce apoptosis.3The results suggest that, Rhizoma polygonati, Morinda officinalis, radix angelicae dahuricae all have antitumor activity when used alone and the combination of three kinds of traditional Chinese medicine, the mechanism may be the role of the cell cycle cells were arrested in S phase and induction of apoptosis. This experiment provides a scientific and theoretical basis for further post-research and clinical applications. |
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