Effect of Inhibition of LncRNA LRRC75A-AS1 on the development of lung cancer cells

ObjectiveTo explore the effect of LncRNA LRRC75A-AS1/miR-22-3p on the proliferation, apoptosis and migration in lung cancer cells.

MethodsThe qRT-PCR method was used to detect the expressions of LRRC75A-AS1 and miR-22-3p in lung cancer tissues and adjacent tissues.Human lung cancer cells A549 were cultured in vitrol, si-NC, si-LRRC75A-AS1, si-LRRC75A-AS1 and anti-miR-NC, si-LRRC75A-AS1 and miR-22-3p inhibitor were transfected into A549 cells.The qRT-PCR method was used to detect the expressions of LRRC75A-AS1 and miR-22-3p in A549 cells.MTT assay and plate clone formation experiment were used to detect cell proliferation and clone formation ability, respectively.Flow cytometry was used to detect the apoptosis rate.Scratch test was used to detect cell migration ability.The dual luciferase reporter experiment was used to detect the targeting relationship between LRRC75A-AS1 and miR-22-3p.Western blotting was used to detect the expression of cleaved-caspase3 protein.

ResultsCompared with adjacent tissues, the expression of LRRC75A-AS1 in lung cancer tissues was increased (1.01±0.13) vs(4.59±0.34)(P < 0.01), and the expression level of miR-22-3p was decreased (1.00±0.12)vs(0.28±0.04)(P < 0.01).Compared with the si-NC group, the cell survival rate of the si-LRRC75A-AS1 group was reduced100%vs(54.45±2.21)%(P < 0.01), the number of clone formation was reduced(121.00±5.10) vs(58.67±2.49)(P < 0.01), the apoptosis rate was increased(8.42±0.34)%vs(25.59±0.80)%(P < 0.01), the protein level of cleaved-caspase3 was increased (0.20±0.01)vs(0.68±0.05)(P < 0.01), and the rate of scratch healing was decreased (66.71±1.43)%vs(32.56±0.95)%(P < 0.01).The dual luciferase report experiment confirmed that LRRC75A-AS1 could act as a competitive endogenous RNA for miR-22-3p.Compared with the si-LRRC75A-AS1+anti-miR-NC group, the cell survival rate of the si-LRRC75A-AS1+miR-22-3p inhibitor group was increased(54.61±2.28)%vs(88.75±3.22)%(P < 0.01), the number of clones was increased (57.67±2.49)vs(106.67±3.68)(P < 0.01), the apoptosis rate was decreased (25.63±0.99)%vs(13.11±0.55)%(P < 0.01), the scratch healing rate was increased (32.52±0.98)%vs(55.30±1.18)%(P < 0.01), and the protein level of cleaved-caspase3 was decreased(0.67±0.06)vs(0.30±0.03)(P < 0.01).

ConclusionsInhibiting the expression of LRRC75A-AS1 may reduce the proliferation, migration and clone formation ability of lung cancer cells by up-regulating miR-22-3p, which can induce cell apoptosis.