The Study Of Gene Expression Spectrum Of Shenqi Fuzheng Injection Role In The Mouse Model Of Liver Metastasis Of Colon Cancer

1 ObjectiveIn hibit the tu mor of the liver metastasis of colon ca ncer miceby va rious interfere progra mmes. The gene expression spectrum ofmouse ca ncer cell was detected by gene chip aftertreatment.A nalyses the spectrum by bioinformatics and find thein herent genet ic cha nges mecha nism of Shenqi Fu zheng Injection indecreasing tu mor and atten uating of small tu mor Rats. Provide thela boratory data for the stud y of function ta rgets of Shenqi Fu zhengInjection.2 Methods2.1 Construct mice model of liver metastasis of colon cancer tumor-bearingSelect BA LB/c mice, esta blish the experimental model by cuttingspleens of mouse after being injected th ree-generation C26 micecolon ca ncer tu mor cells.2.2 Packet,DeliveryMouse were Weighed 24 hou rs after being inoculated , and weredivided into fou r grou ps radomly, na mely , Shenqi Fu zheng Injectiongrou p, 5 - Fu grou p, Shenqi Fu zheng Injection +5- Fu grou p, thecontrol grou p. Begin Intra peritoneal injection ad ministration atesta blish ment model for 5 da ys: 5- Fu injection 0.02 ml (0.5mg),once every other da y, a total of th ree times; Shenqi Fu zhengInjection(Concentrated) 0.15 ml (eq uivalent to 160 mg of Healthmedicine), Overnight a total of five times.2.3 Observation IndicatorsMouse were weighted every da y after being inoculated tu mor.Observe the activities, appetite and the development of tu mor ofmouse. Bu mor- bea ring mouse were killed at the twelfth and fifth da yafter model was esta blished. Tu mor tissue been reserved to sta nd by by freezing with liq uid nitrogen after been removed quickly. Theprepa ration of nucleic acid , detection of chips, analyses ofbioinformatics, verification of the detected results of chips byqua ntitative RT- PCR. After analysis, differential genic data wasentried into the net of Ca pitalBio Corporation and analysis thefunction of differential genes with molecula r annotation system.Cluster analysis and visualization the data to obtain the Clusteranalysis figu re. Finally, it's mecha nism was explained bysea rching gene function.2.4 Technology Road map Detection of gene chip design:4 treatment grou p, two points in time, and repeat th ree timestotalling 7×3 = 21 chips3 ResultsThere are 123 up- regulation different genes in Shenqi grou p andthe control group after lim ma softwa re analysis, and 52 genes amongthem could be sea rched by logined to GeneBa nk, including: A2lp,Anxa7, Atf1, Bbc3, BRIX _MOUSE, Cd h16, Dbr1, Dgkb, Dnajc9, Gluld1,Golgb1, Gp49a , Il13, Itpa , K280_ MOUSE,Laf4, Lrrfip1, Lu , Mospd4,Mre11a , Mst1, N4B3_ MOUSE, NAH7_ MOUSE, Nr1i3, Olf r16, Olf r23,Olf r410, Pitpn m, Prod h, Q8BJ46, Q8C218, Q8C4M4, Q8CB92, Q8CBG9,Q8CG80, Q8K3S6, Q8V HZ3, Q9CS61, Q9CV Q8, Q9CW26, Rad51a p1, Rds,Rnf30, Sca rf2, Sec15l2, Slc9a3, Sprr2a , Tgif , Tmc3, Tmem9, Tpst1,Vps16; the re is one dow n- regulation gene which could n't besea rched in GeneBa nk.52 up- regulation genes were clustered analysis and visualprocessing which function has been described , we ca n see that this52 genes were aggregated into one grou p in Shenqi Grou p and theObserver Grou p, there was significa nt difference between the twogrou ps.30 up- regulation different genes were obtained th rough lim masoftwa re analysis in Shenqi +5- Fu grou p and 5- Fu grou p, and 52genes among them could be sea rched by logined to GeneBa nk,including: Cacna1a, Csn k1e, D14Ertd231e, D4Ertd478e, Fln29, Hic1,K280_ MOUSE, Laf4, Mospd4, Mu p1, Olf r803, Q8C5Y3, Q8C9Y8, Sf3a3;dow n a different gene, log GeneBa nk not sea rch the gene.Beca use of a small number, no, it will not. There is one dow nregulationgene which could n't be sea rched in GeneBa nk. We did n'tma ke cluster analysis for small genes number and there was nostatistical significa nce. 4 ConclusionsColon ca ncer is a com mon ca ncer, liver is the com mon site ofmetastases of colon ca ncer. there are 15% to 25% of the patientsha ve occu rred in the liver metastases when the diagnosis areconfirmed , 20% to 35% of patients with metastatic lesions appea redonly in the liver, 50%～70% ad va nced colon ca ncer patients withliver metastases. Colon ca ncer patients with liver metastases ofcolon ca ncer is the leading ca use of death , therefore, thetreatment of liver metastasis of colon ca ncer is more im portant inclinic. That this stud y used mice liver metastasis of colon ca ncermodel is more close to clinic. Th rough screening differences genesin the treatment grou ps, fou nd that Shenqi Fu zheng Injectionapplied in liver metastasis of colon ca ncer tu mors in mice couldind uce gene express ion in tu mor to occu r a series of cha nges, theoverw helming majority of their function were the up- regulatedexpression.For the differences in genetic screening in Shenqi Fu zhengInjection grou p and the control grou p, we thin k that apoptosisrelatedgene Bbc3 and possessed a function in sta ble ch romosomegene Anxa seven are most noteworth y.It is likely that Bbc3 has a promotion function to allapoptosis signals associated with tu mor which mediated by wildty pe P53. Apoptosis mediated by wild ty pe P53 in huma n colonca ncer cell line could be in hibited by removing Bbc3. In thisstud y, com pa re d with the model grou p, the expression of Bbc3 inliver metastases of Shenqi Grou p was significa ntly increased , itsuggested that Shenqi Fu zheng Injection has a role of againsttu mor and it's mecha nism might related with up relating Bbc3 gene ,therefore, im proving the activity of wild-type P53 gene,promoting the apoptosis tu mor cells.Annexins protein is a kind of protein fa mily which is regulatedby Ca 2 +, is able to com bine with the mem bra ne phospholipid withnegative cha rge and in volved in a series of mem bra ne biologicalactivities related with Ca 2+ . The low expression of Anxa 7 couldlead to multiple signal path wa y interrupted , including tu morsu ppressor genes, DN A repair genes and apoptosis- related genes. Allthese could ca use genomic insta bility. That the expression of Anxa7 in model was signifi ca ntly increased suggested that Shenqi Fu zheng Injection might th rough up regulating Anxa 7 gene andsta bilizing ch romosome to against tu mor.