ABSTRACT
BACKGROUND:Long intergenic non-protein coding RNA 00707(LINC00707)and microRNA-423-5p(miR-423-5p)are both involved in the occurrence and development of osteoarthritis.Starbase predicts that LINC00707 and miR-423-5p have complementary sequences,but whether LINC00707 and miR-423-5p interact to regulate the progress of osteoarthritis still needs further research. OBJECTIVE:To investigate whether LINC00707 targets miR-423-5p to affect chondrocyte injury and inflammatory factor secretion in osteoarthritis. METHODS:Articular chondrocytes were divided into eight groups:(1)blank control group was given no treatment;(2)interleukin(IL)-1β group was cultured with 10 ng/mL IL-1β for 48 hours;(3)IL-1β+si-NC group was transfected with si-NC and then treated with 10 ng/mL IL-1β for 48 hours;(4)IL-1β+si-LINC00707 group was transfected with si-LINC00707 and then treated with 10 ng/mL IL-1β for 48 hours;(5)IL-1β+miR-NC group was transfected with miR-NC and then treated with 10 ng/mL IL-1β for 48 hours;(6)IL-1β+miR-423-5p group was transfected with miR-423-5p mimic and then treated with 10 ng/mL IL-1β for 48 hours;(7)IL-1β+si-LINC00707+anti-miR-NC group was co-transfected with si-LINC00707 and anti-miR-NC and then treated with 10 ng/mL IL-1β for 48 hours;(8)IL-1β+si-LINC00707+anti-miR-423-5p group was co-transfected with si-LINC00707 and anti-miR-423-5p and then treated with 10 ng/mL IL-1β for 48 hours.Relevant tests were performed at the end of the intervention. RESULTS AND CONCLUSION:Compared with the blank control group,the mRNA expression of LINC00707,apoptosis rate,protein expression of C-caspase3 and C-caspase9,and levels of tumor necrosis factor-α and IL-6 in articular chondrocytes were increased in the IL-1β group,while there was a decrease in miR-423-5p expression and IL-10 level(P<0.05).Compared with the IL-1β group,the mRNA expression of LINC00707,apoptosis rate,protein expression of C-caspase3 and C-caspase9,and levels of tumor necrosis factor-α and IL-6 in articular chondrocytes were decreased in the IL-1β+si-LINC00707 group,while miR-423-5p expression and IL-10 level increased(P<0.05).Compared with the IL-1β+miR-NC group,the protein expression of C-caspase3 and C-caspase9 and levels of tumor necrosis factor-α and IL-6 in articular chondrocytes were decreased in the IL-1β+miR-423-5p group,while miR-423-5p expression and IL-10 level increased(P<0.05).Compared with the IL-1β+si-LINC00707+anti-miR-NC group,apoptosis rate,protein expression of C-caspase3 and C-caspase9,and levels of tumor necrosis factor-α and IL-6 in articular chondrocytes were increased in the IL-1β+si-LINC00707+anti-miR-423-5p group,while miR-423-5p expression and IL-10 level decreased(P<0.05).To conclude,inhibiting LINC00707 by targeting miR-423-5p can reduce IL-1β-induced apoptosis and inflammation in articular chondrocytes.
ABSTRACT
Aim To explore the effect of LINC00707 on the proliferation, migration and inflammatory factors of human vascular smooth muscle cells induced by oixidized low-density lipoprotein (ox-LDL) and its possible mechanism. Methods ox-LDL was used to induce human vascular smooth muscle cells (HVSMCs) to establish an atherosclerotic cell model. si-NC, siLINC00707, miR-NC, miR-30c-5p mimic were trans-fected into HVSMCs, and then 100 mg • L~ ox-LDL was added to the cells. si-LINC00707 and anti-miRNC, or si-LINC00707 and miR-30c-5p Inhibitor were co-transfected into HVSMCs and then treated with 100 mg • L ox-LDL. qRT-PCR was used to detect the expression of LINC00707 and miR-30c-5p. CCK-8 and Transwell test were used to detect cell proliferation and migration. ELISA was used to detect the levels of IL-6, TNF-a, and IL-10. The dual-luciferase reporter experiment was used to detect the targeting relationship between LINC00707 and miR-30c-5p. Western blot was used to detect the protein expression of E-cadherin and N-cadherin. Results The expression of LINC00707 in HVSMCs induced by ox-LDL increased (P <0. 05), while the expression of miR-30c-5p decreased (P < 0. 05). After transfection with siLINC00707 or miR-30c-5p mimic, cell viability, the protein level of N-cadherin, the levels of IL-6 and TNF-a decreased (P < 0. 05), and the number of im grating cells decreased (P<0. 05), while the protein level of E-coadherin and the level of IL-10 increased (P <0. 05). LINC00707 could target miR-30c-5p. After co-transfection with si-LINC00707 and miR-30c-5p inhibitor, cell survival rate, the protein level of N-cadherin, the levels of IL-6 and TNF-α increased (P < 0.05), and the number of migrating cells increased (P <0. 05), while the protein level of E-cadherin and the level of IL-10 decreased (P < 0. 05). Conclusion Down-regulation of the expression of LINC00707 could inhibit the proliferation, migration and inflammation of human vascular smooth muscle cells induced by ox-LDL by promoting the expression of miR-30c-5p.