The Research Of The Apoptosis Mechanism In Breast Cancer With Neoadjuvant Chemotherapy Program TEC

Objective:To discuss the TEC program neoadjuvantchemotheroapy in breast cancer cell apoptosis mechanism.Methods:Twopatients with breast cancer neoadjuvant admitted in general surgery ofthe second hospital of the Jilin University from2008to2011wereselected as the experimental subjects.They were definitely diagnosed asinvasive breast cancer by needle core biopsy and their specimens beforechemotherapy were collectted.After four cycles of TEC neoadjuvanttherapy, surgical treatment was adoptted and the tissue samples afterchemotherapy were collectted respectively marked as A1, A2, B1, B2.The specimens of tissue homogenate were used to make up RNA and thetotal RNA was purified parallel inspection.The fluorescent probe should bepurified after the cRNA probe was prepared.Then cRNA should befragmented to produce the chips.We used these gene chips for dataanalysis and verified the consequences of the gene chips by PCRquantitatively in time. Agilent Software and Rotor-Gene Real-TimeAnalysis Software6.0software were applicated for dataanalysis.Results:First:The reports of the quantitative and qualitativeanalysis of the total RNA extracted: the result of electrophoresis was thatthe ratio of28s’ stripe brightness and18s’ was close to2:1and there was no degradation of RNA，which was in line with the requirements of theMicroarray experiments;Second:The result of the hybridization: Byanalyzing microarray hybridization signal，we found that to A sample，2977differential genes were selected out by comparing A1and A2, ofwhich1,643genes were upregulated,1334genes weredownregulated.To B sample,the result was7170differential genes(4264genes upregulated and2906genes downregulated);Third:The analysis ofthe genes’ differential expression: ccording to the genetic screeningmethodology mentioned in the analysis, after we used Agilent’s softwareby the U.S. to Analysze and select data，these probes expresseddifferently were uploaded to NetAFFX Analysis Center of the Agilentwebsite.Then we retrieved the annotation information of thecorresponding probes, including gene title, gene symbol, Gene Ontology,pathway information,and so on. The information should be transformedinto excel documents.Via sorting and classifying the probe information byusing the sorting and searching function of excel,we finally got the cleargenetic names of corresponding probes. We found that IL1-β genes wereupregulated and IL1-α genes lowered.Its associated signal transductionpathways included MAPK signaling pathway.We got2977different probesfor subsequent analysis of biological information by analyzing the differentgene expression which was obtained via comparing the signals of the microarray expression between the experimental and control groups in Agroup. We did gene function analysis based on the different probe,thenwe got43significantly upregulated GO-Term and24significantlydownregulated GO-Term. we analyzed the different signaling pathwaybased on the probes of the difference,23pathways were significantlyupregulated and17pathways were significantly downregul-ated.In GroupB,, we did a differential gene expression analysis about the value of thesignal on the expression microarray of the experimental group and thecontrol group,and achieved7170differential probes for the follow-upanalysis of the biological information.We analyzed the function of genesby the probes of the difference of gene function, we knew that12GO-Terms were significantly upregulated and32GO-Termsdownregulated. we analyzed the difference signaling pathway, by theprobes of the difference of gene function,23pathways were significantlyupregulated and17pathways were significantly down-regulated. Finally,we named the shared genes of the significant difference between groupsA and B as the intersection of genes.We did analysis about the genefunction of the intersection gene,53GO-Terms were significantlyupregulated and27GO-Terms were significantly downregulated. Weachieved the result that there were39Pathways significantly upregulatedand17Pathways significantly downregulated, by the probe based on thedifference signal pathway analysis. Conclusion:The activation of MAPK pathway participates in and launches the apoptosis of the breast cancercells induced by different stimulation.The gene expression level of theIL-1β is up regulation, which ultimately may induce cell apoptosisby activating the JNK and p38MAPK signaling pathway. The geneexpression level of the IL-1α is down regulation, which ultimately mayinduce cell apoptosis by activating the JNK and p38MAPK signalingpathway or inhibit tumor cell proliferation by regulating tumor celldifferentiation and proliferation.