| ObjectiveIn this study,the effect on papillary carcinoma of thyroid cancer,metastasis and invasion of PTC,and the effect of EMT of PTC cells were studied by screening the LncRNA-LUCAT with the highest upregulation in PTC tissue,PTC patient blood and a variety of PTC cell books,and further by knocking down LncRNA-LUCAT in thyroid cancer cell lines.MethodsFrom April 2020 to October 2021,20 PTC tissue samples and adjacent normal tissue samples（within 2cm）obtained in the first Department of general surgery of the Affiliated Central Hospital of Shenyang Medical College,as well as venous blood samples of patients and normal people were selected.All patients（age range 23-76years;mean age 45 years）did not receive any preoperative treatment before thyroidectomy,including radiotherapy,chemotherapy and left thyroxine.It is not accompanied by other malignant tumors（gastric cancer,liver cancer,breast cancer,ovarian cancer,prostate cancer,pancreatic cancer,etc.）or other thyroid inflammatory diseases.All tissue samples were frozen in liquid nitrogen immediately after operation,and their storage conditions were-80 ℃ until further RNA extraction.Through the detection of blood and tissue samples of PTC patients,the expression of lncrnas was detected.Five lncrnas with high expression were screened and verified by RT-q PCR.The lncrna lucat with the highest expression was screened.Then,the expression level of lncrna was detected in many cell lines（thyroid cancer cell lines TPC-1,K1 and ihh-4and human thyroid epithelial cell line nthy-ori3-1）.Lncrna lucat in thyroid cancer cell line TPC-1 was knocked down by intracellular sh RNA.The proliferation of thyroid cancer cell line TPC-1 was further detected by CCK-8 experiment and clone formation experiment,and compared with the negative control sh NC.The apoptosis of thyroid cancer cell line TPC-1 was further detected by flow cytometry and compared with the negative control sh NC.The invasive ability of SH NC group and sh lucat group was detected by Transwell experiment,The migration ability of cells in SH NC group and sh lucat group was detected by scratch test,and the changes of epithelial mesenchymal transformation related proteins in SH NC group and sh lucat group were further detected by Western blot.Further summarize the experimental data and draw the experimental conclusion.The ethics committee of the Affiliated Hospital of Shenyang Medical College approved the above research plan,and all patients expressed informed consent and signed.Fresh samples were sequenced and analyzed by Kangcheng biological company.Results1.The second-generation sequencing results showed that a variety of lncrnas were differentially expressed in PTC tissues and blood of PTC patients: lncrna og,lncrna-w42,lncrna heih,lncrna-h19,lncrna lucat.The expression was up-regulated in PTC tissue and blood of PTC patients;Only lncrna lucat had the highest relative expression level in PTC tissue and blood of PTC patients（P < 0.05）.2.Compared with the human thyroid epithelial cell line nthy-ori3-1,the expression of lncrna lucat was increased in three different PTC cell lines（P < 0.05）.3.Compared with SH NC group,the proliferation of TPC-1 cells in SH lucat group decreased significantly at 24 h,48h and 72 h.After 48 h of cell culture in different groups,the number of cell clones in SH lucat group decreased significantly compared with SH NC group（P < 0.05）.4.Compared with the control group,the apoptosis rate increased after silencing lncrna lucat（P < 0.05）.5.The number of invasive cells in SH NC transfection group was 243.33 ± 24.12,while the number of invasive cells in SH lucat group was 143.34 ± 15.44（P < 0.05）.6.The repair rates of SH NC group and sh lucat group were 45.125 ± 6.543 and35.332 ± 4.453 respectively（P < 0.05）.7.Compared with SH NC group,the expression of E-cadherin and N-cadherin increased and the expression of vimentin decreased in SH lucat group（P < 0.05）.Conclusions1.The expression of lncrna lucat was up-regulated in thyroid cancer tissues and blood;2.Knockdown of lncrna lucat can inhibit the proliferation,migration and invasion of thyroid papillary cancer cells and epithelial mesenchymal transformation,and promote the apoptosis of thyroid cancer cells.3.Lncrna lucat plays the role of oncogene in thyroid papillary carcinoma and may become a potential biomarker for accurate diagnosis and treatment of thyroid papillary carcinoma. |
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