【摘要】 该文旨在探讨长链非编码RNA KCNQ1OT1调节微小RNA(miR)-148a-3p对弥漫大B细胞淋巴瘤(DLBCL)细胞增殖、侵袭、多柔比星(DOX)耐药的影响。实时荧光定量聚合酶链反应(qRT-PCR)检测DLBCL组织及细胞(OCI-LY1)中LncRNA KCNQ1OT1和miR-148a-3p水平;将OCILy1细胞分为对照组(Ctrl)、sh-NC组、sh-K1组、sh-K1+miR-In-NC组、sh-K1+miR-148a-3p-In组;EdU法和细胞克隆形成实验检测各组OCI-Ly1细胞增殖情况; Transwell实验检测细胞侵袭情况;Western blot检测OCI-Ly1细胞中MMP-2、MMP-9和Ki67蛋白表达水平;筛选DOX耐药细胞OCILy1/DOX20,并采用CCK-8法测定各组细胞存活率;双荧光素酶实验测定miR-148a-3p与LncRNA KCNQ1OT1的靶向关系。DLBCL组织或OCI-LY1细胞中LncRNA KCNQ1OT1水平较高, miR-148a-3p水平较低(P<0.05); LncRNA KCNQ1OT1沉默后OCI-LY1细胞EdU阳性率、集落形成数、细胞侵袭数、LncRNA KCNQ1OT1水平以及MMP-2、MMP-9和Ki67蛋白表达水平降低, miR-148a-3p水平升高(P<0.05);进一步抑制miR-148a-3p表达后细胞EdU阳性率、集落形成数、细胞侵袭数、LncRNA KCNQ1OT1水平以及MMP-2、MMP-9和Ki67蛋白表达水平升高, miR-148a-3p水平降低(P<0.05)。在DOX浓度为10～80 nmol/L时, KCNQ1OT1沉默后OCI-Ly1/DOX20细胞存活率降低(P<0.05),进一步抑制miR-148a-3p表达后细胞存活率升高(P<0.05)。KCNQ1OT1与miR-148a-3p间存在靶向关系(P<0.05)。LncRNA KCNQ1OT1下调可能通过调控miR-148a-3p,抑制人DLBCL细胞增殖和侵袭,减弱细胞DOX耐药性。更多还原

【Abstract】 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 KCNQ1OT1 knockdown, 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.更多还原