The Effects Of LCMR1 Antisense Oligodeoxyneucleotides On Biological Behavior Of Highly Metastatic Large-cell Lung Cancer Cell Line-95D And Preliminary Function Study Of LCMR1

Objective Lung Cancer Metastasis Related protein 1 (LCMR1) was a new gene cloned from human large-cell lung cancer 95D cells with highly metastatic abilities and 95C cells with lowly metastatic abilities through differential display reverse transcript PCR(DD-PCR). To explore biological function and mechanism of LCMR1, this study investigated the effects of LCMR1 antisense oligodeoxyneucleotides(ASODN) on biological characteristics of highly metastatic large-cell lung cancer cell line-95D, as well as analyzed the differential gene expression profiles between 95D cells treated by LCMR1 antisense Oligodeoxyneucleotides and the untreated group. Methods (1) LCMR1 antisense Oligodeoxyneucleotides were artificially synthesized and transfected into human highly metastatic large-cell lung cancer cell line-95D. we used RT-PCR to detect the expression levels of LCMR1 mRNA of LCMRJASODN- treated 95D cells and the control groups. Cell growth rate was assayed by MTT, while the ability of cell invasion and mobility were observed by transwell chambers with or without reconstituted matrigel. Colony-forming efficiency was measured by colony formation on double layer soft agarose, cell cycle distribution and apoptotic rate were analyzed by flow cytometry. At the same time, the number of apoptotic cells were examined by invert fluorescence microscope. (2) We compared gene expression profiles of 95D cells betweenLCMR1ASODN-treated group and control group by using high throughput microarray including 4,000 human cDNAs. These findings might provide functional clues of LCMRl.Results (1) After 24 hours treatment of 0.6umol/L ICMKMSODN, the expression of LCMRlmKNA of 95D cells was inhibited to 25%. Compared with the control groups, the growth rate and proliferation ability of ASODN-treated group were markedly reduced, colony-forming efficiency, cell invasion and mobility were decreased. Flow cytometry analysis showed that the percentage of cells in Go/Gi phase was increased, and fluorescence microscope indicated the number of apoptotic cells of ASODN-treated group was obviously increasd. (2) According to the results of hybridization to microarray, the total of 999 differentially expressed genes were screened out. Among the 999 genes, most of them were involved in the physiological processes such as signal transduction, metabolism, protein translation and synthesis. The genes with similar changes of expression were mainly related to regulation of signal transduction of cell proliferation, apoptosis and cell cycle.Conclusions (1) LCMR1 ASODN could significantly reduce the expression of LCMRl mRNAof 95D cells, suppress cell proliferation, inhibit invasion and mobility and induce cell apoptosis. We presumed that gene LCMR1 was closely related to malignant growth of tumor. And also, the results suggested that LCMR1 may play an important role in occurrence and progress of tumor so that it may become a molecular target of gene treatment of lung cancer. (2) LCMR1 may contribute to regulate cell growth differentiation, cell cycle and apoptosis through a signal transduction path which play a key role in malignant growth of rumor. Its precise mechanisms are worthy of further study.