The Study Of MTA1Function In Small Cell Lung Cancer

Objective: Research the biological effects of MTA1in small cell lung cancer. Method:Immunohistochemistry, Western-blot and PCR techniques were used to test, small cell lungcancer-normal lung tissue microarrays, small cell lung cancer cell line H446, MTA1transgenic mice of MTA1expression, through MTA1-siRNA transfected small-cell lungcancer cell lines H446, observe the changes in the malignant behavior of tumor growth,adhesion, apoptosis, cell cycle, and other aspects of invasion and metastasis. Results: Smallcell lung cancer cell lines expressing high levels of MTA1and using MTA1si-RNA reducedits mRNA and protein expression, and verificated results by PCR and western blot.(1) MTA1down-regulation suppressed lung cancer cell line H446proliferation.(2) Down regulation ofMTA1expression impaired the adhesion and invasion ability in SCLC cell line.(3) MTA1silencing slowed down SCLC cells movement in the wound healing assay.(4) MTA1inducessurvival signals in SCLC cell line and down-regulated of MTA1could increased theapoptosis.(5) MTA1high expression in resected SCLC specimens and MTA1transgenicmouse lung tissue, and down-regulated of MTA1silencing on stem cell marker geneexpression. Conclusion: MTA1as a key regulatory factor plays an important biological role insmall cell lung cancer. MTA1involved in the regulation of cancer stem cells, and has asignificant impact in tumor proliferation, invasion, metastasis and apoptosis. Objective: To construct metastasis-associated tumor gene1(MTA1) stably-over-expressingmouse model by inserting the full human MTA1CDS to the genome of C57BL/6J mouse.Method: MTA1gene was cloned form human with RT-PCR, The cloned MTA1gene wasthen inserted eukaryotic expression vector pcDNA3.1to construct of pcDNA3.1-MTA1carrier. The transgenic mouse were produced by microinjection method and the genotype wasdetected by PCR with specific primers. The expression profiles of MTA1in body tissues wereillustrated by Western-blot and immunohistochemical. Results: Three hundred mousezygotes were treated with transgenic vectors DNA by microinjection and then transplantedinto ten artificial pregnant female mice. Ten of these mice were gravid. The transplantationrate was100%.A total of80F0mice were obtained, of which9were carrier MTA1genescreened by PCR. The positive rate of transgenic mice was11.25%(9/80). Four lines ofMTA1transgenic mouse were established. MTA1in transgenic mouse with tissue-specificdistribution and the expression levels of the MTA1gene in brain, liver, lung and colon fromtransgenic mouse were higher than those form control mice. But, kidney and skeletal muscleshowed no difference. Conclusion: Brain, liver, lung and colon of stably-over-expressingMTA1transgenic mouse were established successful. To further MTA1research has laid agood research model.