| Malignant tumor is one of the three killers of human health.Because of its fast growth,fast development and its invasion and metastasis features,it hazard human life. So,earlier diagnosis and valid treatment are critical to malignant tumors.In the 1~st part of my paper,we detected the different mRNA level of ERGIC-53 between leukemia patients and healthy volunteers,between different clilnical stages in CML,and also between drug resistance cell lines and its parental cells.ERGIC-53 may become a novel marker of leukemia for earlier diagnosis,and it may also helps to assess different clinical stages of CML;Integrin avβ3 is expressed on the surface of most malignant tumor cells but not on normal endothelial cell,so it is a good target fot treatment of malignant tumors.In the second part of my paper,we did a lot research on its novel small molecular inhibitor IH1062(3,5-dichloro-phennylbiguanide) on its induction of anoikis and inhibiting of metastasis of integrin avβ3-positive malignant tumors in vivo and in vitro.1 Expression of ERGIC-53 in leukemia.We detected the ERGIC-53 mRNA level by PCR of several pairs of drug-resistant cell lines and their parental cells:K562/A02 and K562,K562/G01 and K562, MCF-7/ADR and MCF-7,and HL60/ADR and HL60.We found that the drug,resistant cell lines K562/A02,K562/G01,and MCF-7/ADR expressed higher level of ERGIC-53 to there parental cells:1.56 folds,1.58 golds and 1.80 folds(p＜0.05),respectively. Meanwhile the drug-resistant cell line HL60/ADR and its parental cell line HL60 had no significant difference in ERGIC-53.The difference and identical of the four pairs of cell lines is:Pgp participates the mechanism of drug resistance in K562/A02,K562/G01,and MCF-7/ADR,while not in HL60/ADR.So we speculate that the cycling protein ERGIC-53 maybe has some relationship with the transport of PgP.Realtime PCR was done with mononuclearcell from 104 leukemia patients and 11 healthy volunteers to detect mRNA level of ERGIC-53.We found that the ERGIC-53 mRNA level of leukemia patients was 142.91±15.42 folds of healthy volunteers(p＜0.05);In CML patients, ERGIC-53 mRNA level was related to the clinical stages:ERGIC-53 of the blast stage/acute stage(314.28±34.73) was higher than that of the chronic stage(66.66±6.43), and the latter was higher than that of the chronic stage(15.23±1.10),p＜0.05.So, ERGIC-53 mRNA level detection is important to clinical of leukemia.It may become a novel marker of leukemia for earlier diagnosis,and it may also helps to assess different clinical stages of CML for better treatment.2 Anoikis induction and metastasis inhibiting of melanoma cells by IH1062 in vivo and in vitro.Integrin avβ3 is a cell adhesion molecule,which is important for the attachment of cells to ECM,responsible for signaling between the cells and the environment,and participates in the regulation of many physiological and pathological processes.IH1062 is a representative compound of a novel type of small molecule inhibitor of integrin avβ3, which were discovered by us recently.In this study,we investigated the anoikis induction of M21 cells by IH1062 the related signaling pathways.Our results demonstrated that IH1062 significantly enhanced anoikis of M21 ceils attached to vitronectin.Meanwhile, the activation of caspase-3,-8,and -9 increased.Moreover,the activation of FAK mediated by vitronectin was inhibited,the protein level of apoptosis-related proteins Survivin and the ratio of Bcl2/Bax decreased.We established a pulmonary metastasis model of melenoma in nude mice.Cells were transplanted to nude mice via the tail vein. Mice were sacrificed before spontaneous death,or on the 50th day after transplantation. The white metastatic foci on the surface of the lung were counted after fixed in Bouin's Buffer.The lung and other organs were sent for pathological biopsy.Results showed transplantation of 1×10~6,2×10~6,and 5×10~6 M21 cells made no difference in metastatic foci on lung surface:83.92±8.04,70.43±6.13,88.35±12.02 respectively.The pulmonary metastasis cells had higher activity of MMP-2(2.76 folds),higher mRNA level of integrin av andβ3 subunit(1.80 and 3.00 fold,respectively),and also less doubling time,compared with the parental M21 cells.So,pulmonary metastasis cells may had more metastasis potential.1×10~6 melanoma cells and 50 days were needed for the model establishment.We then used the pulmonary metastasis model to investigate the anti-metastasis effect of IH1062 in vivo.Metastatic foci of negative control group,0.6 mg/kg group,1.2 mg/kg group,and 2.4mg/kg group were respectively 138.67±26.35, 53.00±4.58,34.00±4.04,and 4.67±2.01.Differences were statistically significance. Pathological check in lungs support the does-dependent effect of IH1062 in inhibiting metastasis.So integrin avβ3 inhibitor IH1062 showed anoikis induction and metastasis inhibiting effects on integrin avβ3-positive malignant tumors. |
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