ESTABLISHMENT OF BONE METASTASIS MODEL OF BREAST CANCER AND EFFECT OF KP-10 ON MIGRATION AND OSTEOCLAST DIFFERENTIATION OF BREAST CANCER CELL LINE BT474

Fei Li¹, Cheng Wang², Binbin Cui³, Xingxia Hu^{3, *}

¹The Fifth Department of Surgery, Changning County Hospital of Traditional Chinese Medicine, Yibin 644300, Sichuan Province, China - ²The Fourth Department of Internal Medicine, Changning County Hospital of Traditional Chinese Medicine, Yibin 644300, Sichuan Province, China - ³Department of Breast and Thyroid Diseases, Taizhou Hospital, Linhai 317000, Zhejiang Province, China

Objective: To analyze the effect of Kp-10 on the migration and osteoclast differentiation of breast cancer cell line BT474. 

Methods: Sixteen clean grade female rats were selected. After anesthesia, the solution prepared by breast cancer BT474 cells was injected between the second and third ribs of mice to establish the bone metastasis model of breast cancer. BT474 cells were stimulated with different concentrations of Kp-10 to observe the number of cell migration. Bone marrow mononuclear cells were isolated and cultured from the femur and tibia. Osteoclast differentiation was induced by breast cancer conditioned medium in vitro. The number of osteoclasts was detected by trap staining. RT-PCR was used to detect the effect of KP-10 on the expression of coordinated genes in bone metastasis of breast cancer. 

Results: After treated with 0.01, 0.05 and 0.1nm concentrations of Kp-10, the migration number of BT474 cells increased gradually with the increase of Kp-10 concentration, which was significantly higher than that of the control group without Kp-10 (P<0.05). The migration number of BT474 cells reached the peak at the concentration of 0.05 nm. The number and area of osteoclasts were observed and counted by trap staining under a microscope. The results showed that the area of osteoclasts was significantly higher than that of the control group (0nm). With the increase of Kp-10 concentration, the number of osteoclasts increased significantly (P<0.05). When the concentration of Kp-10 was 0.05nm, the promoting effect of KP-10 on osteoclast differentiation was the most obvious. Compared with the control group (0nm), the expression of IL-11, CTGF, OPN, MMP1 and RANKL in the treatment of 0, 0.05, 0.1, 0.5 and 1nm concentrations of Kp-10 significantly increased the expression of IL-11, CTGF, OPN, MMP1, RANKL in the control group (0 nm) (P<0.05). 

Conclusion: Kp-10 can significantly promote the migration of breast cancer cells BT474 and osteoclast differentiation induced by breast cancer cells, and the related mechanism may be achieved by promoting the expression of breast cancer synergistic genes.