Gene Expression Of Chemokines In Human Carcinoma Cell Lines With Different Metastatic Potential

Object i ve To get the biological behaviors of breast cancer cell and understand the metastasis mechanism of tumor and Establish a good study model-nude mice model of human breast cancer cell line MDA-MB-435 with different metastatic capabilities that is more suitable for clinical and experimental study. To learn the role of chemokines which are excreted by tumor cells and its adjusting mechanisms of tumor metastasis, screen for a new target of clinical immunotherapy of cancer. Methods Establishment of human breast cancer cell line with different spontaneous pulmonary metastatic capabilities through in vivo selection. Gain human breast cancer cell lines MDA-MB-435 and human hepatocellular carcinoma cell lines MHCC97 with different metastatic potential without any stimulation. Analyse the mRNA expression of four chemokines with RT-PCR. Resu I ts We get new cell lines MDA-MB-435 w, MDA-MB-435F1 x MDA-MB-435F2 and MDA-MB-435HM with different metastatic capability. The following parameter were studied: morphology, karyotype, flow cytometry, migration, the expression of MMP and chemokines. Hypotriploid karyotype: 135w's main range of chromosome is 55-62, 435HM's is 50-52. 4 weeks after injection, the lung metastatic rate was 100% of 435HM; howener, 435W's was less 40%. Flow cytometry revealed cells in S phase for 435W/435HM was 40.34%/55.46%. In vitro invasion assay demonstrated that the cells that penetrated artificial basement membrane was 54?.5 cells per high power field for the former and 117 + 11 cells for the latter. RT-PCR: Human breast cancer cell lines MDA-MB-435 and hepatocellular carcinoma cell lines MHCC97 also can constitutely express all of four chemokines. The amount of every factor is different of two different cell lines. The same trends of chemokines expression are CCL5, CXCL8, XCL1 and CX3CL1. Conclusion New cell lines have been established, which is characterized by different metastasis capabilities. It provides a new model for study of breast cancer metastasis because they have similar geneticbackground. Different tumor cells lines MDA-MB-435 and MHCC97 can constitutely express four chemokines mRNA, with different pattern reflecting metastatic potential at least in partly. It suggests that chemokines with different configuration features may involve in the regulation of the tumor cell's growth and metastasis. It may be necessary to clarify the significance of chemokine production and their regulatory mechanisms.