| Gastric cancer is the most common malignant tumor of alimentary tract and the mortality rate accounts the first in total carcinomas. Radical surgical resection, of which the relapse and metastasis are common, is the main therapeutic method for gastric cancer by now and a lot of patient die from relapse and metastasis. Invasion and metastasis are important biological feature behaviour of malignant tumor. The motility and migration ability of tumor cell plays a key role in saturation and metastasis, and the effective migration behaviour depend on the regulation of intracellular and extracellular regulators to cytoskeleton mechanism, so the tumor research of anti-metastasis medicine, which take cytoskeleton for cure target becomes the new hot-spots. D-Limonene is purified monoterpene from tangerine peel, which has prevention and cure effects on chemo-induced breast cancer, liver cancer, lung cancer, gastric cancer, colon cancer and pancreatic cancer in rodent animal. With the poisonless, extensive and edible features, d-limonene has important research value. The anticancer mechanisms of d-limonene include: induce the expression of metabolic enzyme to detoxicate the carcinogen; induce the apoptosis; redifferentiation of cancer cells and(or) inhibit the isopropylizasion of protein transferase for cell growth and inhibit tumor tiny lymphangiogenesis. But,with respect to inducing reorganization of actin cytoskeleton and inhibiting motility and migration ability on the effect of d-limonene has never been reported now. In our research, we provide a new anti-metastasis theoretical basis for the clinical application of d-limonene through observing reorganization of actin cytoskeleton of gastric cancer cell line SGC-7901 treated with different concentrations of d-limonene and researching the fascin protein which is a F-actin-cross-linking protein.The research includes four parts: Part oneThe study of effects of d-limonene on proliferation and invasive capability in vitro of gastric cancer cell line SGC-7901.Objective: To research the effects of d-limonene on in vitro proliferation and invasive capability of the gastric cancer cell line SGC- 7901.Methods: MTT chromatometry was used to detect the toxic effects on gastric cancer cell treated with different concentrations of d-limonene and define the experimental dosage. colony formatiom were used to estimate the effect of proliferation capability on gastric cancer cell treated with different concentrations of d-limonene. Transwell chambers with reconstituted matrigel were used to estimate the effect of invasive capability in vitro on gastric cancer cell treated with different concentrations of d-limonene.Results: D-limonene, in a dose dependent manner when the dosage of d-limonene is above 0.25mmol/L, has significantly toxic effect on the gastric cancer cell SGC-7901. The proliferation and invasion capability of the gastric cancer cell were prohibited by d-limonene with its growth intacted dosage. different concentrations(0.25mmol/L,0.125mmol/L, 0.0625mmol/L) d-Limonene in the colony formatiom assay, the inhibitition rate were 72.9%,33.3%,6.9%, and 57.6%,45.3%,32.1% from Tanswell assay, respectively.Conclusions: D-limonene has toxic effect on SGC-7901 and significantly inhibite growth in vitro capability of the gastric cancer cell in a dose dependent manner. With its growth intacted dosage, d-limonene significantly inhibite in vitro proliferation and invasive capability of the gastric cancer cell also in a dose dependent manner. Part two The study of effects of d-limonene on reorganization of actin cytoskeleton and F-actin-cross-linking protein fascin of gastric cancer cell line SGC-7901.Objective: To investigate the on reorganization of actin cytoskeleton and the relationship between F-actin and fascin.Methods: To observe the effect of d-Limonene on reorganization of actin cytoskeleton of gastric cancer cell under confocal laser scanning microscopy by using FITC-phalloidin as fluorescent probe. We detected the expression of fascin in the tissues of gastric carcinoma with immunohisto- chemical stain. Western-blot method was used to detect the expression fascin protein of the gastric cancer cell SGC-7901 treated with different concentrations of d-limonene.Results: d-Limonene can induce depolymerization and reorganization of actin cytoskeleton and the changes of arrangement and distribution of microfilame obviously. With immunohistochemical stain, fascin protein was positive in 23 cases among the 58 gastric carcinoma and it correlated with pathological WHO; classification, TNM staging, serosal invasion,lymphatic metastasis. Western-blot results showed the quantity of fascin protein was declined of gastric cancer cell SGC-7901 treated with d-limonene and dose dependent.Conclusions: The expression of fascin is correlated with the distant metastasis capability of gastric carcinoma. D-Limonene can prohibite invasive and metastasis capability of the gastric cancer cell through inducing reorganization of actin cytoskeleton, fascin protein may mediate these process.
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