Huang Yan, Xu Yong-Qiang, Feng Si-Yin, Zhang Xiang, Ni Jiang-Dong
Department of Orthopaedics, The Second Xiangya Hospital of Central South University, Changsha, Hunan Province, People's Republic of China.
Department of Orthopaedics, Hunan Provincial People's Hospital, Changsha, Hunan Province, People's Republic of China.
Cancer Manag Res. 2020 Jun 15;12:4531-4540. doi: 10.2147/CMAR.S248964. eCollection 2020.
This study aimed to investigate the effect of long non-coding TDRG1 on proliferation and migration of osteosarcoma cells through PI3K/AKT signaling pathway.
Altogether 87 cases of osteosarcoma tissues and adjacent tissues were collected, and osteosarcoma cells and osteoblasts were purchased. The expression of LncRNA TDRG1 in tissues and cells was detected by RT-PCR. Si-NC, si-TDRG1, and Sh-TDRG1 were transfected into osteosarcoma cells. L740Y-P (activator of PI3K/AKT pathway) and LY294002 (inhibitor of PI3k/AKT pathway) were used to interfere with PI3k/Akt signaling pathway in osteosarcoma cells. qRT-PCR was used to detect the expression of TDRG1 in osteosarcoma tissues and cells. WB was used to detect the expression of p-PI3K, p-AKT, N-cadherin, E-Cadherin, vimentin, Bax, Caspase-3, and Bcl-2 in cells. CCK-8, Transwell and cell scratch tests were used to detect cell proliferation, invasion and migration, and flow cytometry was used to detect cell apoptosis.
TDRG1 was highly expressed in osteosarcoma, and the levels of p-PI3K and p-AKT were also up-regulated. Cell experiments showed that inhibiting the expression of TDRG1 could inhibit the proliferation, invasion, migration and EMT of osteosarcoma cells, promote the apoptosis of cells, and up-regulating the expression of TDRG1 could promote the proliferation, invasion, migration and EMT of osteosarcoma cells and inhibit the apoptosis of cells. The 740Y-P intervention could reverse the inhibition of Si-TDRG1 on osteosarcoma cell proliferation, invasion, migration and EMT and the promotion of cell apoptosis. LY294002 intervention could reverse the promotion of Sh-TDRG1 on osteosarcoma cell proliferation, invasion, migration and EMT and the inhibition of cell apoptosis.
TDRG1 is highly expressed in osteosarcoma tissue. Silencing the expression of osteosarcoma can inhibit the proliferation, invasion, migration and EMT of osteosarcoma cells by inhibiting PI3K/AKT signaling pathway, which may be a new target for diagnosis and treatment of osteosarcoma.
本研究旨在探讨长链非编码RNA TDRG1通过PI3K/AKT信号通路对骨肉瘤细胞增殖和迁移的影响。
收集87例骨肉瘤组织及癌旁组织，购买骨肉瘤细胞和成骨细胞。采用RT-PCR检测组织和细胞中LncRNA TDRG1的表达。将Si-NC、si-TDRG1和Sh-TDRG1转染至骨肉瘤细胞中。使用L740Y-P（PI3K/AKT通路激活剂）和LY294002（PI3K/AKT通路抑制剂）干扰骨肉瘤细胞中的PI3K/Akt信号通路。采用qRT-PCR检测骨肉瘤组织和细胞中TDRG1的表达。使用WB检测细胞中p-PI3K、p-AKT、N-钙黏蛋白、E-钙黏蛋白、波形蛋白、Bax、Caspase-3和Bcl-2的表达。采用CCK-8、Transwell和细胞划痕试验检测细胞增殖、侵袭和迁移能力，采用流式细胞术检测细胞凋亡情况。
TDRG1在骨肉瘤中高表达，p-PI3K和p-AKT水平也上调。细胞实验表明，抑制TDRG1表达可抑制骨肉瘤细胞的增殖、侵袭、迁移和上皮间质转化（EMT），促进细胞凋亡；上调TDRG1表达可促进骨肉瘤细胞的增殖、侵袭、迁移和EMT，抑制细胞凋亡。740Y-P干预可逆转Si-TDRG1对骨肉瘤细胞增殖、侵袭、迁移和EMT的抑制作用以及对细胞凋亡的促进作用。LY294002干预可逆转Sh-TDRG1对骨肉瘤细胞增殖、侵袭、迁移和EMT的促进作用以及对细胞凋亡的抑制作用。
TDRG1在骨肉瘤组织中高表达。沉默骨肉瘤中TDRG1的表达可通过抑制PI3K/AKT信号通路抑制骨肉瘤细胞的增殖、侵袭、迁移和EMT，这可能是骨肉瘤诊断和治疗的新靶点。
