Studying The Significance Of The DcR3 Gene Expression In Lung Cancer Cell Lines

Objective Studying the significance of the DcR3 gene expressionin lung cancer cell lines.Methods I choose several different lung cancer cell lines for myexperiment, first ,I use 1640-culture medium to cultivate them; second,I use trizol to split cells and obtain the total RNA, then translate theRNA into cDNA ,which as the template for PCR, at the same time I usetwo primers, respectively for DcR3 and β-actin , to synthesize genefragments. After that the products was stained by electrophoresis,thenrunning on 1.0% agarose gel with ethidium bromide ,the products gelimages were obtainted and densities of the products were quantified bygel electrophoresis image analysis system. . by analizing the results bymeans of the Photo Image software ,we can got the information of theexpression of DcR3 gene. the relative expression level of DcR3 genewas calculated as the products of the DcR3 gene divided by that of theβ-actin gene. .The data were handled using CS2000. In thisexperiment , the marker is DL2000, by which I can quantify the gene.Every sample was measured three times. The formula is : the quantityof mRNA =the absorbency of the DcR3 gene/the absorbency ofβ-actingene. The negative results were repeated until I got the reasonableresult. I reclaimed the gel and purified them for sequencing.Results There are several bands we can see at the position of 300bpon the gel clearly. The resultes of sequencing confirm it. The lightnessof bands are different, and the sequence from light to dark is small cellcancer, big cell cancer,gland cell cancer,squama cell cancer, furthermore, so did the absorbency got by the Photo Image software.Discussion RT-PCR is a new ,mature and applied method fordetecting the mRNA. Its merit is high, efficiency and sensitivity. Inthis experiment I use RT-PCR to know what is the difference ofexpression of DcR3 gene of the different four lung cancer cell lines. Ifind that the four cell lines express the DcR3 gene, and the quantity ofgene increase along with the extent of the malignance.I not only detect the expression of DcR3 that is identical to Pitti butalso know the the quantity of gene increase along with the extent of themalignance. The more malignant is the tumor ,the more gene can beobtained , which is the same to the results of S. Hosoe's experiment.In his experiment ,he observed 90 cases who had lung cancer, hefound that the quantity of gene increase along with the extent of themalignance. The higher is the phase,the more is the gene, but in lungcancer ,the relationshio between the different histopathology and theexpression of DcR3 gene is not reported. In my study, in the four celllines, the highest expression of DcR3 gene is the small cell cancer andthe quantity of gene increase along with the extent of the malignance ,and there is no difference in the NSCLC .According to the results wecan continue to study the expression of the DcR3 gene of the differenttumors , the the different tumors in the different stage and before orafter treatment.. by considering the change of DcR3 gene and theclinical parameters we can evaluate the diagnosis, prognosis and thechoosing of drugs. Further more by using the moloclonal antibody ofDcR3, We can block out or debase the expression of DcR3 gene ,andcure the disease from the level of gene.Conclusion the four cell lines express the DcR3 gene, and thequantity of gene increase along with the extent of the malignance. Inthe four cell lines, the highest expression of DcR3 gene is the smallcell cancer ,and there is no difference in the NSCLC .