ABSTRACT
@#[Abstract] Objective: To investigate the effects of long non-coding RNA (lncRNA) taurine up-regulated gene 1 (TUG1) on the proliferation, apoptosis, and epithelial-mesenchymal transition (EMT) of papillary thyroid carcinoma (PTC) cells and its mechanism. Methods: Real-time fluorescent quantitative PCR (qPCR) was used to detect the expression of lncRNA TUG1 in 35 pairs of PTC tissue and corresponding paracancerous tissue specimens that were surgically resected in Tangshan Workers’ Hospital, Hebei Province from May 2019 to April 2021, human PTC cell lines (TPC-1, BHP10-3, K1, SW1736) and human normal thyroid epithelial Nthyori 3-1 cells. TPC-1 cells were cultured in vitro and divided into control group, si-NC group, si-TUG1 group, miR-NC group, miR-524-5p mimic group, si-TUG1+NC inhibitor group, si-TUG1+miR-524-5p inhibitor group, miR-524-5p mimic+pc-DNA group, and miR-524-5p mimic+ pcDNA BRAF (V-raf murine sarcoma viral oncogene homolog B1) group by transfecting corresponding vectors. CCK-8 and FCM methods were used to detect the proliferation and apoptosis of TPC-1 cells, WB method was used to detect the expression of BRAF, proliferating cell nuclear antigen (PCNA), cysteine protease-3 (Caspase-3), E-cadherin, N-cadherin and vimentin in TPC-1 cells, and the dual-luciferase reporter gene experiment was used to verify the targeting relationship between miR-524-5p and lncRNA TUG1 as well as BRAF. Results: lncRNA TUG1 was up-regulated in PTC tissues and cells (P<0.05). Silencing the expression of lncRNA TUG1 or up-regulating the expression of miR-524-5p could significantly inhibit the proliferation and EMT, and promote apoptosis of TPC-1 cells (all P<0.05). Dual-luciferase reporter gene experiment showed that there was a targeting relationship between miR-524-5p and lncRNA TUG1 as well as between miR-524-5p and BRAF (all P<0.05). Silencing the expression of miR-524-5p could reverse the effects of lncRNA TUG1 knockdown on inhibiting proliferation and EMT and promoting apoptosis of TPC-1 cells (all P<0.05), and up-regulation of BRAF expression could reverse the effects of miR-524-5p overexpression on inhibiting proliferation, EMT and promoting apoptosis of TPC-1 cell (all P<0.05). Conclusion: lncRNA TUG1 is up-regulated in PTC tissues and TPC-1 cells. Silencing the expression of lncRNA TUG1 can inhibit proliferation and EMT but promote cell apoptosis of PTC cells by regulating the miR-524-5p/BRAF axis.
ABSTRACT
@#Objective: To investigate the expression of lncRNA01296 in esophageal cancer (EC) tissues and its effect on the proliferation and migration of EC TE-2 cells. Methods:Atotal of 36 pairs of esophageal cancer tissues and corresponding para-cancerous tissues were collected from EC patients admitted to the Department of Thoracic Surgery,Affiliated Hospital of Chengde Medical College from January 2017 to September 2018. The human normal esophageal epithelial (HEEC) cells and human esophageal cancer cell lines ECA109, TE-1 and TE-2 were cultured. qPCR was used to detect the mRNAexpressions of lincRNA01296, SNRPA(small nuclear ribonucleoproteinA) and NGF (nerve growth factor) in EC tissues and cells. Recombinant lentiviral interference vectoror control vector were used to transfect EC cell lines, as sh-lncRNA01296#1,#2 and Mock groups. WB was used to detect the protein expressions of SNRPAand NGF in transfected cells. MTS assay was used to detect cell proliferation, and Transwell assays were used to detect cell invasion and migration of TE-2 cells after transfection. Results: The mRNAexpressions of lncRNA01296, SNRPAand NGF were significantly increased in esophageal cancer tissues and cell lines (all P<0.01), and these expressions in poorly differentiated TE-2 cells were higher than those in highly differentiated ECA109 and TE-1 cells (all P<0.05). The mRNAexpressions of lncRNA01296 and NGF in sh-lncRNA01296#1 and sh-lncRNA01296#2 groups were significantly lower than those in Mock group (all P<0.01), while the mRNAexpression of SNRPAshowed no statistical difference among three groups (P>0.05). The protein expressions of lncRNA01296 and NGF in sh-lncRNA01296#1 and sh-lncRNA01296#2 groups were significantly lower than those in Mock group (all P<0.01). The relative proliferation ability of cells in sh-lncRNA01296#1 and shlncRNA01296#2 groups was significantly lower than that of Mock group at 48 and 72 h after transfection (P<0.05 or P<0.01). The number of invasive cells was (72.0±6.3), (36.6±4.3) and (33.9±3.7) in Mock, sh-lncRNA01296#1 and sh-lncRNA01296#2 groups, respectively; and the number of migrated cells was (85.2±9.9), (47.5±8.1) and (43.8±6.5), respectively, indicating that the numbers of invasive and migrated cells in sh-lncRNA01296#1 and sh-lncRNA01296#2 groups were significantly less than those in Mock group(all P<0.01). Conclusion: lncRNA01296 can up-regulate SNRPAexpression to promote NGF-mediated proliferation and metastasis of EC cells, which may provide new target for the diagnosis and treatment of esophagealcancer.