| Gastric cancer is a common malignant tumor,especially in China,its incidence and mortality ranked second in the country.The clinical treatment of gastric cancer model is mainly to take surgical resection.But a large number of patients with gastric cancer was diagnosed into the progress period.For these patients,usually it has been used systemic chemotherapy,radiotherapy treatment,but due to the existence of serious adverse reactions,the quality of life of patients has not been significantly improved,and the survival rate is still very low.The invasion,metastasis of gastric cancer is the main reason for the low survival rate of cancer patients.The transfer process is also very complex,mainly including tumor cells from the primary site,and gradually infiltration of peripheral tissue and into the blood circulation system and lymphatic circulation system,through the cycle of distant target organs and tissues,the formation of metastases.Gastric cancer invasion,metastasis is a serious threat to human health,therefore,to find a natural effective treatment of malignant tumors is particularly urgent.Elemene is a compound that is isolated from the rhizome of Curcuma sinensis,which is widely used as a medicament for the treatment of various solid tumors.Its anti-tumor mechanism is very complex,including the direct inhibition of killing tumor cells,induced apoptosis of tumor cells differentiation,anti-tumor cell infiltration and metastasis.Its main ingredient β-elemene as a new type of anti-tumor monomer compounds have also shown its unique advantages.It can regulate multiple signaling pathways involved in the phosphorylation of multiple kinases,such as Ras / MARK /ERK and PI3 K / Akt / m TOR.These studies further elucidate the anti-tumor mechanism of β-elemene-induced apoptosis and autophagy of the body.However,the underlying mechanism by which β-elemene is associated with the treatment of gastric cancer is largely unknown.In recent years,lysyl oxidase(LOX)has played a key role in the proliferation,invasion and metastasis of malignant tumor cells.The effect of β-elemene on gastric cancer and its association with extracellular matrix-regulated enzyme LOX needs further study.Objectives:1 To clarify the effect of β-elemene on the proliferation,apoptosis and cycle of human gastric cancer cells.2 To clarify the effect of β-elemene on the metastasis and invasion of human gastric cancer cells.3 To clarify the mechanism of β-elemene on gastric cancer cells.Methods:1 The effect of β-elemene on the survival rate of human gastric cancer cell line was detected by MTS method in human gastric adenocarcinoma cell line BGC823 and SGC7901.There were 9 groups in the negative control group,the positive control group and the experimental group(50,100,200,300,400,600 and 800 μmol / L).BGC-823,SGC7901 were exposed to β-elemene 24 h,MTS method was detected the growth inhibition by OD value.Hoechst 33342 cell staining was used to determine the effect ofβ-elemene on cell apoptosis in gastric cancer cell lines,Flow cytometry was used todetect the effect of different concentrations of β-elemene on cell apoptosis and cell cycle in human gastric cancer cell lines.2 The effects of different concentrations of β-elemene(100,200,400 μmol/L)on the metastasis of human gastric cancer cells were detected by Transwell metastasis assay.Transwell invasion assay was used to detect the effects of different concentrations ofβ-elemene(100,200,400 μmol /L)on the invasion of human gastric cancer cells;3 The effect of β-elemene on the activity of LOX was detected by enzyme-linked immunosorbent assay.The amount of total RNA in the cells was extracted and the expression of LOX was quantitatively analyzed by real-time fluorescence quantitative method.These expressions of LOX,intergrinβ1,FAK,E-cadherin and N-cadherin were assayed by western blot.Results:1 The effect of β-elemene on proliferation and apoptosis of human gastric cancer cell line Human gastric cancer cell lines were expored to β-elemene for 24 hours and observed by light microscope.The morphological changes of gastric cancer cells were observed in the drug intervention group.The cells were gradually shrunk from the original clear,the cells were not closely connected,the apoptotic bodies were seen in the cytoplasm,and a large number of floating cells appeared.The results showed that gastric cancer cells treatment with different concentrations of β-elemene,and in the range of a certain concentration.Hoechst 33342 cell staining showed that the concentration of β-elemene in the treatment of human gastric cancer cells,with the increase in drug concentration,blue clumps,chromatin shrinkage and highlight.And the number of cells is gradually reduced,and the emergence of irregular debris.The results showed that β-elemene could promote the apoptosis of human gastric cancer cells.Flow cytometry analysis showed that the increase of apoptotic rate was related to the drug concentration after treatment of gastric cancer cells with different concentrations of β-elemene.The apoptosis rates of BGC823 were 6.67%,8.73%,9.65% and 17.90%,respectively.The apoptosis rates of human gastric cancer cell line SGC7901 were 8.1%,8.2%,18.7% and 41.9%,respectively.Compared with the control group,the apoptosis rate was significantly higher.The ratio of S phase cells in BGC823 and SGC7901 cells was decreased and the proportion of G2/M phase cells increased after treated with different concentrations ofβ-elemene.2 Effects of β-elemene on the metastasis and invasion of selected human gastric cancer cells2.1 Effect of β-elemene on the metastasis of gastric cancer cells The results of wound healing assay showed that the treatment group could significantly inhibit the scoring distance(P<0.05)after treated with different concentrations of β-elemene in gastric cancer cells.With the increase of drug concentration,the number of BGC823 and SGC7901 cells decreased and the metastasis ability of gastric cancer cells decreased gradually.Transwell metastasis experiments showed that the treatment group of gastric cancer cells treated with different concentrations of β-elemene could significantly inhibit the number of transmembrane cells.2.2 Effects of β-elemene on the invasion of gastric cancer cells Transwell invasion experiments showed that the treatment group of gastric cancer cells treated with different concentrations of β-elemene could significantly inhibit the number of transmembrane cells.of LOX,integrinβ1,FAK,E-cadherin and N-cadherin Treated with different concentrations of β-elemene(100,200,400 and 800 μmol/L)for24 h with SGC7901,compared with the control group,LOX enzyme activity decreased in a dose-dependent manner,and decreased expression of LOX m RNA(P<0.05).Compared with the control group,the expression of LOX,integrinβ1,FAK and N-cadherin in the gastric cancer cells treated with β-elemene(100,200,400 and 800μmol/L)were down-regulated,E-cadherin protein up-regulated.3 Effects of β-elemene on LOX Enzyme Activity,LOX m RNA and the expressionConclusions:These results demonstrate that the proliferation,metastasis and invasion of human gastric cancer cell lines BGC823 and SGC7901 can be inhibit by the treatment ofβ-elemene and it can induce apoptosis of gastric cancer cells.The effect of β-elemene on gastric cancer may be related to reducing LOX enzyme activity,LOX m RNA level,down-regulation of LOX,Integrinβ1 and FAK,and induce epithelial-mesenchymal transition in tumor cells to inhibit tumor metastasis and invasion. |
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