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AIM: To study the effect and potential mechanism of IFN? on tumor angiogenesis. METHODS: Human umbilical cord vascular endothelial cell (HUVEC) was cultured in vitro. The effect of IFN? (10, 100, and 1000 unit?ml -1) on proliferation of HUVEC was detected by MTT assay in vitro. HUVEC pretreated with IFN? (10, 100, and 1000 unit?ml -1) was stained with PI and detected by flow cytometry (FCM). The mRNA expression of apoptosis relative gene Bcl-2 and Bax of HUVEC treated with IFN? (100, and 1 000 unit?ml -1) were detected with semi quantitative reverse transcription- polymerase chain reaction (RT-PCR). IFN? (30, 300, and 3 000 unit?ml -1) on VEGF secretion and expression in human lung carcinoma cell PG in hypoxia were detected by ELISA assay. CONCLUSION: The key attributes of IFN? on tumor angiogenesis possible directly inhibit vascular endothelial cells or indirectly inhibit growth factors secretion and expression in tumor cells.
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AIM: To detect sequence and mutation of K-ras oncogene in tissue and stool DNA of patients with colorectal cancer in order to provide a method of noninvasive and simple colorectal cancer diagnosis. METHODS: DNA was separated and purified from colorectal cancer tissue or stool of patient with colorectal cancer, then the K-ras gene was amplified by PCR and PCR products were cloned, the K-ras gene was sequenced, and the mutation was identified. The expression of color/colorectal cancer antigen was inspected by immunohistochemical technique. Stool sample of patient with colorectal cancer was detected with enzyme-linked immunosorbent assay (ELISA). RESULTS: K-ras gene sequence of the stool was completely same as that of the tissue of the patient;K-ras mutation was detected in one case. There was relativity between the mutation of K-ras gene and the pathology type of colorectal cancer and the expression level of colorectal cancer antigen in stool sample. CONCLUSION: It is feasible that colorectal tumors can be detected by a noninvasive method based on the molecular pathogenesis of the disease. Detecting K-ras gene mutations of stool DNA can provide bases for the screening, early detection, and prognosis to patients with colorectal cancer.
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Objective:To find out the effects of Ganoderma lucidum polysaccharides peptide (Gl-PP) on the invasion of the human lung carcinoma cell (PG cell). Methods: PG cells were pretreated with different concentration Gl-PP in vitro, using cell proliferation assay, cell migration assay, adhesion assay, zymography and RT-PCR, then the effects of Gl-PP on proliferation, motility, adhesion and MMP-9 activity and mRNA expression of PG cells were investigated in vitro. Results: Gl-PP did not directly inhibit PG cell proliferation in vitro. However, pretreated with Gl-PP, PG cells motility was inhibited significantly. PG cells adhesion was also inhibited. The activity of MMP-9 was inhibited in a dose-dependent manner, and the inhibited ratio was 41.53% at a dose of 100 mg/L Gl-PP. The mRNA expression of MMP-9 of pretreated PG cells was inhibited. Conclusion: Gl-PP could suppress invasion of human lung carcinoma cells in vitro.
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AIM: To investigate the influences of nimodipine on HCT cells proliferation and explore the mechanism. METHODS: HCT cells were treated with different concentrations of nimodipine, and its proliferation was inspected by MTT assay. The apex areas of sub diploid were measured by Flow Cytometry and the DNA ladder were found by agarose gel electrophoresis. The characteristic changes in morphology were observed under the light microscopy. The cellular distribution and concentration of calcium were studied by using the laser confocus scanning microscopy. RESULTS: The growth of HCT cells was inhibited by different concentrations of nimodipine. Data from Flow Cytometry showed the apex areas of sub diploid enlarge in the drug treating group, suggesting that the number of apoptosis cells increased in dose dependent manner. Gel electrophoresis displayed DNA cleavage pattern typical of apoptosis: DNA ladder in high dose nimodipine treated HCT cells. The light microscopy presented cellular morphological changes: cell membrane blebs, the cytoplasm and nuclear chromatin condensation. The concentration of cytosolic free calcium increased when treated with nimodipine showed by the laser confocus scanning microscopy. CONCLUSION: Nimodipine can cause inhibition of the cell proliferation of HCT cells. The mechanism of inhibition might involve the cell apoptosis.