Impacts of LncRNA KCNQ1OT1 on Proliferation, Invasion, and Doxorubicin Resistance of Diffuse Large B-Cell Lymphoma Cells by Targeting miR-148a-3p
GU Zheyun, TAO Jian, WANG Ling*
This study aims to investigate the impacts of LncRNA KCNQ1OT1 on proliferation, invasion, and DOX (doxorubicin) resistance of DLBCL (diffuse large B-cell lymphoma) cells by regulating miR(microRNA)-148a-3p. qRT-PCR (quantitative real-time polymerase chain reaction) was used to measure the levels
of LncRNA KCNQ1OT1 and miR-148a-3p in DLBCL tissues and OCI-Ly1 cells. OCI-Ly1 cells were assigned into Ctrl (control) group, sh-NC group, sh-K1 group, sh-K1+miR-In-NC group, and sh-K1+miR-148a-3p-In group. EdU method and cell clone formation assay were used to detect the proliferation of OCI-Ly1 cells in each group. Transwell experiment was used to detect cell invasion. Western blot was used to detect the expression levels of MMP-2, MMP-9, and Ki67 proteins in OCI-Ly1 cells. DOX resistant cells OCI-Ly1/DOX20 were screened, and CCK-8 method was used to determine the survival rate of cells in each group. Dual luciferase assay was used to determine the targeting relationship between miR-148a-3p and LncRNA KCNQ1OT1. LncRNA KCNQ1OT1 in DLBCL tissue or OCI-LY1 cells was higher, and miR-148a-3p was lower (P<0.05). After LncRNA KCNQ1OT1 knockdown, the EdU positivity rate, colony formation number, cell invasion number, LncRNA KCNQ1OT1 level, MMP-2, MMP-9, and Ki67 protein expression levels of OCI-LY1 cells in the sh-K1 group were prominently lower, while miR-148a-3p were prominently higher (P<0.05). After further inhibition of miR-148a-3p expression, the EdU positivity rate, colony formation number, cell invasion number, LncRNA KCNQ1OT1 level, MMP-2, MMP-9, and Ki67 protein expression levels of OCI-LY1 cells in the sh-K1+miR-148a-3p-In group were prominently higher, while miR-148a-3p were prominently lower (P<0.05). When the DOX concentration was between 10-80 nmol/L, the survival rate of OCI-Ly1/DOX20 cells in the sh-K1 group was prominently lower after LncRNA KCNQ1OT1knockdown, and the survival rate of OCI-Ly1/DOX20 cells in the sh-K1+miR-148a-3p-In group was prominently higher after further inhibition of miR-148a-3p expression (P<0.05). There was a targeting relationship between KCNQ1OT1 and miR-148a-3p (P<0.05). The downregulation of LncRNA KCNQ1OT1 may inhibit the proliferation and invasion of human DLBCL cells and weaken cell DOX resistance by regulating miR-148a-3p.
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