| Objective:Lung cancer is one of the most common malignant tumors,and its morbidity and mortality rank first in the country.Lung cancer has many pathological types,among which lung adenocarcinoma is one of the most common pathological types.Although certain results have been achieved in the treatment of lung cancer,due to the high mutation of its gene sequence,the phenomenon of resistance to chemoradiotherapy is also increasing It is widespread,so finding new therapeutic targets is the current research hotspot.Long non-coding RNA?LncRNA?is a type of newly discovered RNA that does not encode proteins and is longer than 200bp.In recent years,research on it has made certain progress,and it has been found in many tumors Abnormal expression in cells.It can play the role of a regulatory factor,participate in tumor progression,and promote tumor proliferation and transformation.Metabolic reprogramming is one of the ten major characteristics of tumors.In recent years,targeted energy metabolism therapy has become a new research hotspot and has been used to a certain extent.Therefore,this study mainly explores long-chain non-coding RNA related to urothelial carcinoma1 on the glucose metabolism of lung adenocarcinoma,whether UCA1 can be used as a new therapeutic and screening target,and further explore its impact on the biological behavior of lung adenocarcinoma.Methods:In this study,lung adenocarcinoma A549 and H1299 cell lines and human normal bronchial epithelial cells HBE were studied.The expression of UCA1 in A549,H1299 and HBE was first detected by qRT-PCR.The A549 and H1299 cell lines were then transfected.Both cell lines were transiently transfected with short hairpin RNA?sh-RNA?and plasmid?UCA1?to up-regulate UCA1,and then qRT-PCR was used to detect UCA1 in A549 and Transfection efficiency of H1299.The transfected A549 and H1299 cell lines were used to detect the expression changes of glucose metabolism?GLUT1,HK2,PKM2,LDHA?by qRT-PCR and Western Blot,to observe the effect of up-regulation of UCA1 on glucose metabolism index;In the lactic acid experiment,the lactic acid kit was used to detect the change of lactic acid content in the medium,and the obtained values were statistically analyzed by t test.By FDG uptake assay,18F-FDG uptake changes,to filter out the normal A549 and uptake peak H1299 cells,the CPM changes of A549 transfected and H1299 cell line into the detection of the peak,calculating FDG uptake by the formula Finally,the change of cell proliferation rate was detected by EDU proliferation experiment.The changes of invasion and migration ability were detected by Transwell and scratch test.One-way analysis of variance was used among multiple groups of data obtained,and two-sample independent t test was used for comparison between the two groups.Results:1.The expression of UCA1 in A549,H1299 and HBE cell lines was detected by qRT-PCR.The results showed that UCA1 was highly expressed in A549 and H1299 cell lines compared with HBE,and the highest expression was found in A549 cell line.Significance?P<0.001?.2.The two cell lines were transfected with qRT-PCR and Western Blot,respectively,to detect changes in glucose metabolism at the RNA and protein levels.The qRT-PCR results of A549 and H1299 cell lines showed that the glucose metabolism in the sh-UCA1 group compared with the NC group.The indicators were all decreased,while the UCA1 group and the Vector group showed a significant increase in glucose metabolism at the RNA level,and the difference was statistically significant,P<0.01.Western Blot results showed that the expression levels of GLUT1,HK2,PKM2 and LDHA in sh-UCA1 group were lower than those in NC group,while the expression level in overexpression group was significantly higher than that in negative control group?P<0.05?.3.After transfection,EDU assay was used to detect the proliferation of the two cell lines.The results showed that the cell proliferation rate increased significantly after over-expression of UCA1,while the cell proliferation rate decreased after silencing UCA1,the difference was statistically significant,P<0.01.4.After transfection of A549 cells,the cell invasion numbers of sh-UCA1 group,NC group,UCA1 group and Vector group were 17±4,65±7,99±7 and 53±5,respectively?each field of view?,H1299 cells were transfected.After infection,the number of cell invasion in sh-UCA1 group,NC group,UCA1 group and Vector group was 55±7,100±6,163±16,98±8?each field of view?,and the difference was statistically different,P<0.001.The cells were transfected and scratched.The scratches were compared at 0 hours and 24hours.The results showed that the cell migration rate decreased significantly after down-regulating UCA1,and the migration rate increased after up-regulating UCA1.Therefore,down-regulating the expression of UCA1 can inhibit cell invasion.And migration capabilities.5.The lactic acid kit was used to detect the lactic acid content of the sugar metabolite in the medium.The results showed that the sh-UCA1 group?1.06±0.06;0.86±0.08?was compared with the NC group?2.16±0.15;1.53±0.11?in the two cell lines.The lactic acid content decreased significantly,while the over-expressed UCA1group?2.95±0.2;2.73±0.15?had higher lactic acid content than the Vector group?1.89±0.09;1.66±0.14?,and the difference was statistically significant.6.After the experiment using FDG uptake UCA1 transfection,both cell lines FDG uptake rate changes found after reduction of A549 and H1299 cells UCA1 uptake rate decreased?2.12±0.13;2.8±0.25?,whereas overexpression of FDG uptake rate after UCA1 The increase was?3.55±0.24;5.69±0.18?,the difference was statistically significant,P<0.05.Conclusion:In summary,the long-chain non-coding RNA UCA1 can act as a regulator and act as a molecular marker and therapeutic target.UCA1 plays a role in promoting tumor progression in lung adenocarcinoma,which can promote the energy metabolism of tumors,increase the ability of tumor cells to proliferate,invade and migrate.Therefore,UCA1 can be used as a new therapeutic target to inhibit the progression of tumors. |
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