| ObjectiveThyroid papillary carcinoma(Papillary Thyroid Carcinoma,PTC)is the most common thyroid malignant tumor,clinical treatment has limitations and side effects and other shortcomings,so it is particularly important to find cheap,less side effects and efficient drugs.In this study,TPC-1 cell line was subcutaneously inoculated into Balb/c nude mice to replicate the transplanted tumor model of thyroid papillary carcinoma,to observe the inhibitory effect of ursolic acid on the transplanted tumor of thyroid papillary carcinoma,and to explore the effect of ursolic acid on Wnt/β-catenin signal pathway,so as to further reveal the molecular mechanism of ursolic acid inhibiting the proliferation of thyroid papillary carcinoma.It is expected to provide new research ideas and methods for the early treatment of thyroid papillary carcinoma in the future.Method1 Establishment of HPLC fingerprint of Prunella vulgaris from different habitats and determination of ursolic acidThe fingerprints of Prunella vulgaris from 16 batches of different producing areas were established by HPLC,and the content of ursolic acid in Prunella vulgaris from different areas was determined.The quality control of Prunella vulgaris was analyzed,and the Prunella vulgaris with the highest content of ursolic acid was selected.Prunella vulgaris from this area was used as the experimental material for later research.2 Effect of ursolic acid on the construction of subcutaneously transplanted tumor model in nude mice by TPC-1 cell lineBalb/c nude mice were evenly divided into tumorigenesis group,solvent control group,low,medium and high concentration UA groups.Thyroid papillary carcinoma TPC-1 cell line was subcutaneously inoculated into the left forelimb of nude mice to replicate the subcutaneous transplanted tumor model of thyroid papillary carcinoma in nude mice.The tumor-forming group was not treated,the solvent control group was injected intraperitoneally with 200 μL control solvent,and the UA group was intraperitoneally injected with ursolic acid of the same concentration,once a day for 7days,and then observed for 14 days.The volume of transplanted tumor was measured and recorded every 3 days during the experiment.At the end of the experiment,the nude mice were killed,the transplanted tumor was stripped and weighed.The transplanted tumors were divided into groups and embedded in wax slices,and the effect of ursolic acid on the morphology of tumor cells in nude mice was observed by HE staining.To determine whether ursolic acid can inhibit the subcutaneous tumorbearing of TPC-1 cell line in nude mice.3 Mechanism of ursolic acid inhibiting proliferation of thyroid papillary carcinoma through wnt/β-catenin signal pathwayThe expressions of related factors WIF-1,GSK3 β,Cyclin D1,c-myc,Tnks,DKK,SFRP4,JRK,FOXM1,SOX7,PYGO2 m RNA and protein in wnt/β-catenin signal pathway in different groups of transplanted tumors were detected by QRT-PCR and Western blot methods.To explore the possible mechanism of ursolic acid inhibition on thyroid papillary carcinoma.Result1 Establishment of HPLC fingerprint of Prunella vulgaris from different habitats and determination of ursolic acidThe results showed that the similarity between the fingerprints of the 16 batches of Prunella vulgaris samples from different areas and the control fingerprint was 0.883-0.999,indicating that there were some differences in the quality of the 16 batches of Prunella vulgaris samples.Among the 16 producing areas of Prunella vulgaris studied,the content of ursolic acid in Prunella vulgaris from Wenshan Prefecture of Yunnan Province was the highest,so Prunella vulgaris from Wenshan Prefecture of Yunnan Province was selected as the experimental material for later research.2 Effect of ursolic acid on the construction of subcutaneously transplanted tumor model in nude mice by TPC-1 cell lineThe results showed that ursolic acid significantly inhibited the growth of tumorbearing volume compared with tumor-forming group and solvent control group.The results of HE staining of tumor tissue in nude mice showed that the tumor cells in the tumorigenesis group and the solvent control group were arranged in pieces,the size was consistent,the nucleus was intact,the proportion of nucleus and cytoplasm was larger,and more vascular sections could be seen in the tumor tissue,while in the low,middle and high concentration groups of UA,the tumor nucleus became smaller,pyknosis,the proportion of nucleus to cytoplasm became smaller,and even necrosis occurred in some areas,and there were fewer blood vessels in the tumor.3 Mechanism of ursolic acid inhibiting proliferation of thyroid papillary carcinoma through wnt/β-catenin signal pathwayThe results of QRT-PCR and Western blot methods showed that ursolic acid could up-regulate the expression of WIF-1,GSK3 β,SFRP4,DKK,SOX7 m RNA and protein in wnt/β-catenin signal pathway,and down-regulate the expression of β-catenin,Tnks,JRK,FOXM1,PYGO2,Cyclin D1,c-myc m RNA and protein,suggesting that ursolic acid can affect the proliferation and apoptosis of thyroid carcinoma by regulating wnt/β-catenin signal pathway.ConclusionThe transplanted tumor model of nude mice replicated by TPC-1 cell line was used as the experimental object.Ursolic acid inhibited the tumor volume and weight of nude mice after acting on the tumor-bearing model of nude mice.According to QRT-PCR and Westernblot experiments,ursolic acid can inhibit the proliferation of thyroid papillary carcinoma through wnt/β-catenin signal pathway. |
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