| Purpose and background: Lung cancer is one of the most common cancer in theworld, lung cancer can be divided into two types: non-small cell lung cancer (NSCLC)and small cell lung cancer (SCLC), NSCLC (Non small-cell lung cancer, NSCLC)accounts for80%~85%of all cases of lung cancer, non-small cell lung cancer hasbecome the world’s highest morbidity and mortality of malignant tumor, lung cancermortality of malignant tumor mortality both men and women are the first, a seriousthreat to human health In recent years, with the development of molecular biology andpharmacology of comprehensive treatment of NSCLC and individualized therapy hasachieved significant progress, however, due to the emergence of drug-resistant tumor,makes most of the tumor chemotherapy appeared soon after recurrence or metastasis,limiting the many chemotherapy drugs such as paclitaxel antitumor drugs in thetreatment of NSCLC.BFGF is a fibroblast mitogen, a major role in originates from neural ectodermand mesoderm fibroblasts, skeletal muscle cells and bone cells, etc. FGF exist twokinds of receptors: one kind is low affinity receptors, receptor called heparin samples,for heparan sulfate proteoglycan. Another type of affinity for the receptor, is atransmembrane receptor tyrosine protein kinase existence; Different FGF receptorexpression has tissue specificity. The present study confirmed the tumor cell growth, survival, apoptosis and angiogenesis in the process of FGFs FGFRs signalingpathways abnormal activation and participate in the process. Study found that patientswith lung cancer associated with prognosis in patients with bFGF content in serum,serum content increased, the prognosis is poor. Previous research found that bFGFinvolved in pathological and physiological mechanisms of angiogenesis related,speculated that bFGF not only in the occurrence, development, invasion andmetastasis of lung cancer play an important role in the process, and the body’sresistance.Build target bFGF siRNA to bFGF liposomes as the carrier mediated siRNAtransfection A549lung adenocarcinoma cancer cells, using a real-time fluorescentquantitative reverse transcription polymerase chain reaction (reverse transcription-polymerase chain reacti, RT-PCR detection of bFGF siRNA interference by bFGFmRNA content in cells before and after transfection screening by the most significantof siRNA sequences Using protein immunoblot detection technology (western blotanalysis and western blot) detection of bFGF siRNA interference by bFGF proteinlevels in lung adenocarcinoma A549cells before and after the change is determinedby MTT method applied standard colorimetric method to detect bFGF siRNAinterference within the tumor cells before and after the cut of bFGF expression levelrelations with taxol drug susceptibility, to provide theoretical basis for clinicalindividual therapy.Materials and Methods: Used PUBMED search bFGF gene sequences, the NM002006.4, siRNA sequences using artificial synthesis, at the same time, to negativesiRNA in comparison, can the bFGF with Lipofectamine2000liposomes as thecarrier siRNA into transient transfection, lung adenocarcinoma A549cells of bFGFsiRNA interference by lung adenocarcinoma A549cells before and after extraction oftotal RNA and protein, rt-pcr technique was applied to siRNA interference of lungadenocarcinoma A549cells before and after the cut bFGF mRNA for quantitativeanalysis Select high transfection efficiency of siRNA sequences. Reuse screeningsequence of siRNA interference cut lung adenocarcinoma A549cells, steps are thesame as before, Westen blot method detecting interference after the downgrade in lung adenocarcinoma A549cells the expression of bFGF protein level. Determined byMTT colorimetry to detect interference downgrade of taxol drug sensitivity in lungadenocarcinoma A549cells before and after the change. Flow cytometry is used totest cut noise in lung adenocarcinoma A549cells apoptosis and cell cycle changes.Results:RT-pcr results showed that bFGF after siRNA interference by lungadenocarcinoma A549bFGF gene expression level decreased, after which bFGFsiRNA-2gene sequences cut most efficient transfection; Testing results of Westernblot showed siRNA interference-1cut lung adenocarcinoma A549cells after theirbFGF protein expression level of interference significantly lower before cut.Determined by MTT colorimetry to detect lung adenocarcinoma A549cells beforeand after the cut on the taxol drug sensitivity, results show that the interference cellsto taxol inhibition rate than the control group before and after the downgrade wassignificantly higher (P <0.05). Flow cytometry instrument apoptosis detection shows:bFGF siRNA plus paclitaxel group of apoptosis cell percentage control siRNA groupcontrol siRNA plus paclitaxel group bFGF siRNA group increased significantly (P <0.05) in the cell cycle detection show that the control siRNA plus paclitaxel group andbFGF siRNA plus paclitaxel group G2/M phase cells percentage is higher than thecontrol group siRNA and bFGF siRNA group, the difference was statisticallysignificant (P <0.05)Conclusion:Interference by SiRNA can decrease bFGF gene expression level,increase the lung adenocarcinoma A549cells to taxol drug sensitivity, promote taxolinduce apoptosis and inhibit cell growth... |
PDF Full Text Request