Capsaicin Induce The Cisplatin-mediated Osteosarcoma Apoptosis By Inhibiting The Epression And Nuclear-translocation Of Id1

Background:The early transfer and chemotherapy resisitance are the major reasons for the medicial failure of the tumor patients.However,the common chemotherapy drug is weak to reduce the expression of anti-apoptosisi related gene while add more drug only cause the side effect rather than improving the effect.So it is important to find the resistance-related gene and the substance which point the target.Recent years,clinical evidence proved the inhibitor of differentiation had the potential value by comparing the gene and protein in the different tumor staging,the results shown that various kinds of tumor including prostatic cancer,bladder cancer,lung cancer and maglinant glioma had shown higher level of Id1.More evidences shown that both of development of cancers and chemotherapy resistance are connected with the expression of Id family.Hao had proved that the tumor expressed highr level of Id1 is more likely to transfer compared with the lower group while those induce the death.Those research had been proved by Chen while they prove that the maglinent lung tumor cell line show more Id1 expression level compared with benign cell line.those evidences show the connection between Id1 with the grade maglinent.Higher expression level of Id1 would improve the VEGF expression and induce the transfer.On the other hand,growing evidences show that the consistant expression of Id1 may enhance the chemotherapy resisitance,First,knockdown the Id1 expression could effectively reduce the weight of tumor and enhance the effect of cisplatin and doxorubicin.the evidence show the maglinent tumor would be more susceptible if the expression of Id1 was effectively knocked.So,the researchers pointed that Id1 could be the potential target and remove the Id1-related resistance.Osteoscaroma is the most common tumor in the orthopedics,First,it most possibly happened in the tibia,femur and humerus that would influent the movement of the limbs.Surgery alone would accompany with the higher relapse rate,so it is important to use the chemotherapy to reduce the chances.Clinical researchs shown that cisplatin and doxorubirin and cyclophosphamide are the most effective drug.The result show that the chemotherapy increase the 5 year survive rate to 60-70%since the 1970s.Second,the teenagers and olders are the most suspetical people,these trend need the doctors considering the doseage to avoid the side effects.The osteosarcoma is the abnormal osetoblasts cell which weak the maturation,and the Id1 play the important role in the process.evidences show that the more Id1 expression,the higher grade Ennecking the tumor was that proved the connection between the Id1 with the maglinent grade in the ostesarcoma.the Id1 is the downstream of the Bone Morphogenetic protein that involve the control of the necessary gene expression.It had been improved that Id1 involved the cell surivive,proliferation,transfer and chemotherapy resistance by the nuclear translocation.Thus,inhibite Id1 could inhibit the Id1-mediated drug resisitance and improve the effect of clinical process.Our team had proved that that the importance of Id family in the differentiation of the bone,the expression of Id1 is important for the cell proliferation in the early osteoblasts cell.Knockdown the Id1 would influence the BMP-mediated bone induce especially the BMP9.recent year we found that the abnormal differentiation may cause the osteosarcoma.However,even the Id1 is important for the tumor,the cuase of Id and mechanism that Id1 involve are little discuss which make the curation point the Id1 difficult.1.Osteosarcoma express high level of Id1 and induce the cisplatin resistanceObjectiveanalyse the different level of Id1 between osteosarcoma and control group to confirm the higher expression level in osteosacroma.Test the function of cisplatin in three different osteosarcoma cell lines.Identify the advance of knockdown Id1 in cisplatin-mediated chemotherapy resistance.Methods1 Collect the tissue from 12 osteosarcoma patients and the bone tissue from 12 osteoarthritis which were tested by IHC,the results show the distinction of the Id1 expression.2 The Western Blot was used to test the expression change of Id1 after using cisplatin in three osteosarcoma cell lines MG63,143b and HOS.3 IF was used to test the nuclear translocation of Id1 after treating cisplatin in three osteosarcoma cell lines.4 Using virus to construct the knockdown Id1 cell lines MG63-Id1,143b-Id1 and HOS-Id1,the CCK-8 and cell flow technology were used to analyse the function of Id1 in cisplatin-mediated apoptosis.Result1 IHC technology confirmed the higher level of Id1 in the osteosarcoma compared with the control groupand the mesenchyme.2 The high dose of cisplatin could induce the expression of Id1 in three osteosarcoma cell lines.3 The result of IF show that cisplatin induce the nuclear translocation of Id1.4 The result of IF showed that the connection of Id1 and p65 in the nuclear after treating cisplatin.5 The groups of knockdown of Id1 were more sensitive to cisplatin?Conclusionthe higher level of Id1 in the osteosarcoma were specificity.Thus,the substance which knockdown the Id1 expression may have the medicinal prospect in specific usage.Meanwhile,the Knockdown-Id1 group showed more sensitive in cisplatin treatment that prove the conclusion mentioned above.Here,we find that cisplation induce the Id1 nuclear translocation and connection with p65,the result mean that function pathway of Id1 and how the Id1 in inducing the chemotherapy resistance.2.capsaicin reduce the expressionof Id1 by NF-Kb and TGF signal pathway.ObjectiveTo test whether the capsaicin could reduce the Id1 epression of id1 in osteosarcoma and the involved pathway.Method1.Using different dose of capsaicin to treat the osteosarcoma cell lines in different times.2.Using Western Blot and qPCR to clarify the way of capsaicin in reduce the Id1.3.Using Western Blot to explose the potential way that capsaicin in reducing Id1?4.Using inhibitor of NF-kB and TGF signal pathway to clarify the connection between Id1 and capsaicin.Result1.capsaicin could reduce the expression of Id1 protein while induce the transription of Id1.2.the activity of NF-kB induce the Id1 protein.Capsaicin induce the transcription of Id1 by actitiving the TGF signal pathway.ConclusionCapsaicin could reduce the Id1 protein by dose-and time-depedent manner while induce the transcription of Id1.The AKT-IKB-p65 signal pathway is connected with the capsaicin-mediated knockdown of Id1.Capsaicin induce the transcription of Id1 by actitiving the TGF signal pathway rather than BMP signal pathway.Using the specific inhibitor of these pathway confirmed that both AKT-IKB-p65 and TGF signal pathway could inhibit the expression of Id1 which are connected with eath other.3.capcaicn inhibit the cisplatin-mediated Id1 expression and nucleartranslocaion and enhance the cisplatin-mediated apoptosisObjectiveTo clarified the function of that combined both capsaicin and cisplatin in osteosarcoma in vitro and explored the function of capsaicin related change of signal pathway in enhancing the cisplatin-mediated chemotherapy.Methods1 Using the CCK8 test tested the effect of unite of capsaicin and cisplatin2 Using the cell flow tested the effect of unite of capsaicin and cisplatin3 Western blotwas used to analyse the difference of capsaicin and cisplatin in Id1-related mechanism.4 IF was used to clarified whether capcaicin could inhibited the nuclear translocation of Id1.Result1 Combine of capsaicin and cisplatin could effectively reduce the activity of osteosarcoma cell lines.2.Cell flow proved that combine of capsaicin and cisplatin could effectively induce the apoptosis of osteosarcoma cell lines.3.Capsaicin could effectively inhibited the cisplatin-mediated phosphorylation of p65 and Smad 1/5 to reduce the Id14.IF proved that Combine of capsaicin and cisplatin could reduce the nuclear translocation of Id1 which was prove by Western blotConclusionThe combine of capsaicin and cisplatin could effectively enhance the chemotherapy effect in osteosarcoma.We also prove that capsaicin could eliminate the cisplatin-related change of NF-kB and TGF signal pathway and reduce the expression of Id1.The inhibition may also work though the inhibition of nuclear translocation of Id1 in osteosarcoma cell lines.All these finally induce the apoptosis of osteosarcoma by the combine of capsaicin and cisplatin...