Ki67 Antisense Oligodeoxynucleotide On The Inhibition Of Breast Cancer Cell Proliferation, Invasion And Related Mechanisms Of

Part IOBJECTIVE: To investigate the correlation between the expression of Ki67 and the malignant phenotype in human breast cancer cells. METHODS:The expression of Ki67 in three breast caner cell lines was detected by half-quantitative RT-PCR and Western blot assay.MTT was used to detected the proliferation capability.Transwell assay was used to evaluate the capability of migration and invasion in those cell lines in vitro,respectively. RESULTS:The level of transcription and translation of Ki67 was coincident in three breast caner cell lines(P<0.05).Moreover,the capability of proliferation and invasion show the same trend:MDA-MB-435s> MDA-MB-231> MCF-7. CONCLUSION: There was a positive correlation between the expression level of Ki67 and the capability of proliferation and invasion in three of the breast cancer cell lines (r=0.82,0.78).Thus,Ki67 may play multiple roles in the process of breast cancer occurrence and development as an independent factor,and may be a new molecular target for cancer therapy.Part IIOBJECTIVE: To study the inhibition effects of an antisense Ki67 vector on proliferation and invasion of cervical carcinoma cell line MDA-MB-435s,and explore interrelated mechanism. METHODS: The conservative region of Ki67 gene was amplified by RT-PCR and then cloned into pEGM-T vector.After the recombinant plasmid was certified by DNA sequencing,the conservative region of Ki67 gene was inserted into pEGFP vector reversedly. Then the eukaryotic expression vector for Ki67 antisense RNA was constructed and transfected into malignant cervical carcinoma cell line,MDA-MB-435s, by lipofectamine. The expression levels of Ki67 before and afer transfection were detected by RT-PCR and Western blot assay;MTT and Boyden assay was used to detected the capability of proliferation and invasion in vitro,respectively; Flow cytometry analyzed apoptosis. RESULTS: The expression levels of Ki67 mRNA and protein decreased sharply in antisense Ki67 transfected cells. The inhibition rate was respectively. CONCLUSION: The antisense Ki67 vector could inhibit markedly the proliferation,invasion of cervical carcinoma cell, and contributed to apoptosis or silencePart IIIBACKGROUND﹠OBJECTIVE: The purpose of study is to determine the expression of Ki67 and to explore the relationship between Ki67 and lymphatic metastasis in invasive breast ductal cancer. METHODS: Tissue sections from 60 invasive breast ductal cancer patients were examined immunohistochemically to detect the expression of Ki67 in malignant portion and adjacent normal tissue. The correlation between the expression of Ki67 and clinicopathological features was analyzed. RESULTS: Positive Ki67 immunostaining was detected in 51 cases of 60 breast carcinoma(85%). The median scores of Ki67 expression was 3.9 in lymph node metastasis group versus 2 in lymph node negative group (P<0.05). The expression of Ki67 in lymphatic node level I.II and III groups is significantly higher than that in lymph node negative group(P<0.01). CONCLUSION: The high expression of Ki67 is correlated with existence and distance of lymphatic metastasis in breast cancer.