Study On The Expression And Significance Of Rab25and ERK In Osteosarcoma

Osteosarcoma is a malignant tumor that occurs in adolescents. The malignancydegree of it is very high, the prognosis is poor.It can occure metastasis in short time.The development of osteosarcoma is a multistage process complicated with multiplegenes.It is the focus of current research to find critical genes in the tumordevelopment.It is important to block and intervent it.Rab25is located on chromosome1q22, is a newly discovered Ras oncogenefamily subfamily member, and is homologous with Rab11, it is involved in dynamicprocess. Previous studies have found that Rab25expression was higher in ovariancarcinoma and breast carcinoma than that of other organizations cancer. Rab25wasreported that become the treatment point of ovarian cancer and a new treatmentapproach. There are researchs that shew that Rab25was related with osteosarcomaoccurrence and prognosis closely.ERK is a member of MAPK signal transduction system. It is involved in cellgrowth, differentiation, metastasis of the cell and has been found higher expression ina variety of tumors.ObjectiveThe expression of Rab25and ERK mRNA and protein in the osteosarcoma andits adjacent tissues was tested by our study. The relationship between the two genewas explored.The object of this study was to find the osteosarcoma tissues specificcancer genes and specificity strong targets.The study can provide basis for earlydiagnosis and treatment of osteosarcoma.Materials and methods1.Specimens collection: Osteosarcoma and its tumor adjacent tissues were takenfrom the first affiliated hospital of the first affiliated hospital of Henan University, thefirst affiliated hospital of Zhengzhou University and the Huaihe Hospital of HenanUniversity.All patients signed the informed consent. They are13-26years old, in which conventional osteosarcoma18cases, telangiectatic osteosarcoma9cases,3cases of small cell osteosarcoma. The corresponding normal surrounding bone tissuewere taken at the same time.All the tissues were diagnosed by pathology. Collectionof fresh specimens were immediately stored in liquid nitrogen, then transferred to-80℃refrigerator.2.RT--PCR (reverse transcription polymerase chain reaction): RNA wasextracted according to the <molecular clone>. The precipitate air dried and washedwith50μlDEPC ddH20,All the operation was completed in low temperatureenvironment. the The absorbance of RNA was merasured by absorbance and theconcentration was calculated． The same amount of RNA was Taken and operatedaccording to the description of RT Kit．3.Western blot: The protein was extracted according to the <molecular clone>.The gel of proteins was transferred to nitrocellulose membrane.After that，themembrane was incubated with primary antibody,second antibody in turn.DAB kti wasused to stain at last．The hybridization effect was observed and the target proteinlocation was determined.Flashed it with ECL (chemiluminescence method) methodin the dark room finally.4. Half quantitative analysis:β-actin as a reference in Rt-PCR and Western blotand Bioimaging System and GeneTools software were used to analyze the aimbands.The expression level of Rab25and ERK mRNA and protein in cancer tissuesand its matched adjacent tissues were tested by semi-quantitative technique．5. Statistics methods：Data of the experiment was processed by SPSS13.0software．The two groups were compared with t test,the rate was compared using thechi-squared test, significance level for test was P <0.05.Result1.RT-PCR semi quantitative results: The relative expression of Rab25inosteosarcoma tissue was0.5231+0.1331and0.1226+0.0115in the adjacent tissues,the difference was statistically significant (P <0.05). The relative expression of ERKin osteosarcoma tissue was0.4632+0.1237and0.1134+0.0193in the adjacenttissues, the difference had statistical significance (P <0.05). 2.Western blot semi quantitative results: The relative expression of Rab25inosteosarcoma tissue was0.4326+0.1058and0.1024+0.0145in the adjacent tissues,the difference was statistically significant (P <0.05). The relative expression ofERK in osteosarcoma tissue was0.5466+0.1895and0.1256+0.0111in the adjacenttissues, the difference had statistical significance (P <0.05).3. Rab25and ERK mRNA and protein expression differences in differentpathological types of osteosarcoma tissue had no statistical significance (P>0.05).Conclusion1.The expression of Rab25and ERK mRNA in osteosarcoma tissues was higherthan that of the adjacent tissues.2. The expression of Rab25and ERK protein in osteosarcoma tissues was higherthan that of the adjacent tissues.3. Rab25and ERK mRNA and protein expression differences in differentpathological types of osteosarcoma tissue was not statistically significant.