| Part 1 Expression of heat shock protein 70 in breast cancer and its clinical value analysisBreast cancer is the most common malignant tumor in our country, the sixth position in the cause of cancer death, which seriously threatens women’s physical and mental health. With the development of molecular pathology and molecular biology, it has been a great clinical research to improve the early diagnosis, comprehensive treatment and prognosis of breast cancer. At the same time, the research and development of anticancer drugs targeting biological markers related targets, also become the basis of breast cancer and clinical research hot spot, including the role of heat shock proteins in breast cancer. By qualitative and quantitative methods, the expression of heat shock protein 70 in different breast tissues and its relationship with molecular biology were analyzed to investigate the biological indicators of breast cancer metastasis and prognosis, and to provide some experimental evidence for the diagnosis and prognosis of breast cancer.Objective To observe the expression of heat shock protein 70 in breast cancer, atypical hyperplasia, carcinoma adjacent tissues and normal tissues, and analyze its relationship with breast cancer related factors.Methods 160 cases of breast cancer confirmed by surgical resection were collected, and the clinical data were obtained. The expression of HSP70 in different tissues was detected by qualitative and quantitative methods, and the relationship between them was analyzed.Results (1) HSP70 protein was expressed in breast cancer tissues, and the expression of HSP70 was correlated with tumor size, axillary lymph node metastasis, HR and Her-2, but there was no significant correlation between pathological type, menstrual status and histological grade. HSP70 positive group (n=70), no disease progression time was 24 months (95%CI:22.0-26.1), and no disease progression was 33 months (95%CI:29.8-36.2 months), Rank=11.6 P=0.001, Log. Kaplan-Meier analysis showed that the overall survival time was 32 months (95%CI:26.8-37.2 months) in HSP70 positive group (n=70), while the overall survival time was 39 months (n=50), Rank=6.9 Log, P=0.012.(2) The expression of mRNA HSP70 in breast cancer and atypical hyperplasia was significantly higher than that of Real-timePCR (P<0.05), but the expression of HSP70 was not changed significantly (P=0.82). The expression of HSP70mRNA was positively correlated with tumor size, pathological type, histological grade, PR status, Her2 status, and ER expression in axillary lymph nodes (rs=0.792, P<0.001).Conclusion HSP70 has a certain degree of expression in breast cancer tissues, HSP70 positive expression is related to the prognosis of patients with breast cancer.Part 2 Effect of heat shock protein 70 on biological function of breast cancer cell lines in vitroBreast cancer is the most common malignant tumor in China, which seriously threatens women’s physical and mental health. Our previous studies have shown that HSP70 expression is associated with the clinicopathological features of breast cancer, and it is associated with the development and progression of breast cancer, and is an indicator of poor prognosis in breast cancer patients. For further understanding of the mechanism of its role, we apply the gene transcription method to detect the expression of HSP70 gene expression of strains in vitro breast cancer MDAMB231 cell anti proliferation effect and biological function; by Western blot and real-time PCR was used to detect the HSP70mRNA in MDA-MB-231 cells in expression level, and migration ability by MTT assay, Transwen chamber method, microscope to observe and compare the transfection of HSP70mRNA gene in MDA-MB-231 cells line and adhesion ability and morphological change and to further reveal the HSP70 in breast cancer occurrence and development mechanism provides experimental basis.Objective To investigate the effect of heat shock protein 70 gene expression on the biological function of breast cancer cell line in vitroMethods The migration ability of MDA-MB-231 cells to construct a lentiviral vector (HSP70mRNA. Application of gene transcription method to detect HSP70 gene expression effects on the in vitro breast cancer cell line MDA-MB-231 antiproliferative effect and biological function of; by Western blot and real-time PCR was used to detect the HSP70mRNA in MDA-MB-231 cells in expression level, and by MTT method. Transwen chamber method to, loss of cell technology and inverted microscope, to observe and compare the transfection HSP70mRNA gene, adhesion ability and morphological change.Results 1.MDA-MB-231 cells transfected with HSP70mRNA containing cells, the expression of HSP70 in mRNA and protein levels were significantly increased, and the level of HSP70 protein was 2.7 and 3.5 times of the non transfection group.2. The proliferation activity of MDA-MB-231 cells was detected by MTT assay, and the adhesion ability was significantly enhanced (p<0.05); Under the inverted microscope, the cells were transfected into the cells, the growth of 72h was good, and the cells were not significantly reduced, and the positive clones were observed;The apoptosis rate of MDA-MB-231 breast cancer cells in different transfection groups were detected by flow cytometry, and the apoptosis rate was 1.17% after 48h, and the apoptosis rate was 17.81% in 48h group, and 19.80% in the blank group (P<0.05);After transfection of 48h cells, the cells gradually enter the blank area, and then the cell "scratch" of 72h cells after the cells were cultured; The transfected cells with high expression of HSP70mRNA had obvious migration and invasion ability.Conclusion HSP70 gene transfection can promote cell proliferation, invasion and apoptosis in breast cancer MDA-MB-231 cell line, which may be one of the factors that affect the prognosis of patients with breast cancer. |
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