Effects of KCNQ1OT1 Gene Knockout Combined with Bruceine D on Proliferation, Migration, and Invasion of Breast Cancer MDA-MB-231 Cells / 肿瘤防治研究

Objective To explore the effect of KCNQ1OT1 gene knockout combined with bruceine D on the proliferation, migration, and invasion of breast cancer MDA-MB-231 cells. Methods Cell Counting Kit-8, wound healing, and Transwell invasion assay were used to detect the effects of bruceine D and siKCNQ1OT1 on the viability, migration, and invasion of MDA-MB-231 cells. Effect of bruceine D and siKCNQ1OT1 on the expression of KCNQ1OT1 in MDA-MB-231 cells was detected by qRT-PCR. Western blot was used to detect the effect of bruceine D and siKCNQ1OT1 on the expression of EMT-related proteins and CDC42, p-MKK7, MKK7 proteins in MDA-MB-231 cells. Results Bruceine D and siKCNQ1OT1 could significantly inhibit the viability, migration, and invasion of MDA-MB-231 cells, and the inhibitory effect was enhanced when they were combined (all P < 0.05); bruceine D downregulated the expression of KCNQ1OT1 in MDA-MB-231 cells (all P < 0.05); bruceine D combined with siKCNQ1OT1 significantly decreased CDC42, p-MKK7, N-cadherin, and Vimentin expression in MDA-MB-231 cells and increased the expression of E-cadherin (all P < 0.05). Conclusion Bruceine D combined with siKCNQ1OT1 significantly inhibit the proliferation, migration, invasion, and EMT of human breast cancer MDA-MB-231 cells, and its molecular mechanism may be related to the blocking of CDC42/MKK7 signaling pathway.

lncRNA KCNQ1OT1 affects the proliferation and apoptosis of retinal epithelial cells induced by high glucose through miR-19a-3p/TSHZ3 / 国际眼科杂志(Guoji Yanke Zazhi)

@#AIM: To investigate whether long-chain non-coding RNA(lncRNA)KCNQ1OT1 affects the proliferation, apoptosis and oxidative stress of retinal epithelial cells induced by high glucose(HG)through miR-19a-3p/TSHZ3. <p>METHODS: Cell counting kit 8(CCK-8)was used to detect the cell viability of human retinal epithelial cells ARPE-19 stimulated with 5, 15, 45, 135mmol/L HG. The ARPE-19 cells were divided into NC group, 45mmol/L HG group, si-NC+45mmol/L HG group, si-lncRNA KCNQ1OT1+45mmol/L HG group, miR-NC+45mmol/L HG group, miR-19a-3p mimics+45mmol/L HG group, si-con+45mmol/L HG group, si-TSHZ3+45mmol/L HG group, pcDNA+si-lncRNA KCNQ1OT1+45mmol/L HG group, pcDNA-TSHZ3+si-lncRNA KCNQ1OT1+45mmol/L HG group. CCK-8 was used to detect cell viability, qRT-PCR was used to detect the expressions of lncRNA KCNQ1OT1, miR-19a-3p and TSHZ3 mRNA, Western Blot was used to detect TSHZ3, activation-cysteine-containing aspartate proteolytic enzyme 3(Cleaved-caspase-3), B-cell lymphoma/leukemia-2(Bcl-2)related X protein(Bax)protein expressions, and enzyme-linked immunosorbent assay(ELISA)was used to detect the levels of oxidative stress indicators reactive oxygen species(ROS)and malondialdehyde(MDA). The dual luciferase activity was used to detect the targeted binding between lncRNA KCNQ1OT1 and miR-19a-3p, miR-19a-3p and TSHZ3. <p>RESULTS: 15, 45, 135mmol/L HG inhibited the survival rate of ARPE-19 cells, and the subsequent select the HG concentration 45mmol/L with a cell survival rate of about 50%. 45mmol/L HG increased the expression levels of lncRNA KCNQ1OT1, TSHZ3 mRNA, TSHZ3 protein, the apoptosis rate, Cleaved-caspase-3 and Bax protein expressions, ROS and MDA levels in ARPE-19 cells, and reduced cell survival rate and the expression level of miR-19a-3p(<i>P</i><0.05). Low expression of lncRNA KCNQ1OT1, TSHZ3 or high expression of miR-19a-3p improved the survival rate of ARPE-19 cells induced by HG, and reduced apoptosis rate, Cleaved-caspase-3 and Bax protein expressions, ROS and MDA levels(<i>P</i><0.05). lncRNA KCNQ1OT1 targeted miR-19a-3p, miR-19a-3p targeted TSHZ3, and lncRNA KCNQ1OT13 regulated the expression of TSHZ3 through miR-19a-3p. The effect of lncRNA KCNQ1OT1 low expression on the survival rate, apoptosis and oxidative stress of ARPE-19 cells induced by HG was reversed by the overexpression of TSHZ3.<p>CONCLUSION: The low expression of lncRNA KCNQ1OT13 promotes the proliferation of retinal epithelial cells induced by high glucose, and inhibits their apoptosis and oxidative stress through miR-19a-3p/TSHZ3.

Kcnq1ot1 promotes osteogenic differentiation and suppresses osteoclast differentiation / 南方医科大学学报

OBJECTIVE@#To investigate the regulatory role of long non-coding RNA Kcnq1ot1 in osteoclast differentiation, osteogenic differentiation and osteoporosis.@*METHODS@#The expression of lnc-Kcnq1ot1, Bglap, Runx2, Alp, Bsp, Nfatc1, Mmp9, Ctsk and Oscar were detected by real-time quantitative PCR (qRT-PCR) in the femoral bones from mouse models of postmenopausal osteoporosis (ovariectomized mice, @*RESULTS@#The expression of lnc-Kcnq1ot1 was significantly upregulated during osteoblast differentiation but downregulated in the bone tissues of osteoporotic mice and during osteoclast differentiation (@*CONCLUSIONS@#Our data demonstrate that lnc-Kcnq1ot1 promotes osteogenic differentiation and alleviates osteoclast differentiation, suggesting the potential of lnc-Kcnq1ot1 as a therapeutic target against osteoporosis.

lncRNA KCNQ1OT1 reverses the effect of sevoflurane on hepatocellular carcinoma progression via regulating the miR-29a-3p/CBX3 axis

Sevoflurane (SEVO) is widely applied as an anesthetic, which exerts antitumor capacity in various cancers, including hepatocellular carcinoma (HCC). Previous studies indicated that long non-coding RNA KCNQ1 opposite strand/antisense transcript 1 (KCNQ1OT1) was upregulated, while microRNA-29a-3p (miR-29a-3p) was downregulated in HCC. Thus, we aimed to explore the roles of KCNQ1OT1 and miR-29a-3p in HCC cells exposed to SEVO. Cell proliferation, apoptosis, migration, and invasion were assessed by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, flow cytometry, and transwell assays, respectively. The levels of genes were determined by quantitative real-time polymerase chain reaction (qRT-PCR) or western blot. Furthermore, the interaction between miR-29a-3p and KCNQ1OT1 or chromebox protein homolog 3 (CBX3) was predicted by Starbase or Targetscan, and then confirmed by dual-luciferase reporter assay. We found that the levels of KCNQ1OT1 and CBX3 were decreased, while miR-29a-3p was increased in SEVO-treated HCC cells. KCNQ1OT1 overexpression weakened the inhibitory effects of SEVO on HCC cell proliferation, apoptosis, migration, and invasion. Interestingly, KCNQ1OT1 bound to miR-29a-3p, and miR-29a-3p targeted CBX3. KCNQ1OT1 upregulated CBX3 level by repressing miR-29a-3p expression. Furthermore, KCNQ1OT1 exerted tumor promotion in HCC cells via suppressing miR-29a-3p to regulate CBX3 expression. Collectively, our findings demonstrated that KCNQ1OT1 regulated the antitumor effects of SEVO on HCC cells through modulating the miR-29a-3p/CBX3 axis, providing a theoretical basis for the treatment of HCC.

Long non-coding RNA KCNQ1 overlapping transcript 1 promotes the progression of esophageal squamous cell carcinoma by adsorbing microRNA-133b

OBJECTIVE: The long non-coding RNA (lncRNA) KCNQ1 overlapping transcript 1 (KCNQ1OT1) exerts vital regulatory functions in diverse tumors. However, the biological function of KCNQ1OT1 in esophageal squamous cell carcinoma (ESCC) remains unclear. METHODS: KCNQ1OT1 expression was detected in ESCC tissues using quantitative real-time polymerase chain reaction (qRT-PCR). Cell proliferation, apoptosis, migration, and invasion were detected by the CCK-8 assay, EdU assay, flow cytometry analysis, and Transwell experiments, respectively. Bioinformatics analysis, luciferase reporter experiments, and RNA immunoprecipitation assays were used to predict and validate the regulatory relationships between KCNQ1OT1, microRNA-133b (miR-133b) and epidermal growth factor receptor (EGFR). RESULTS: KCNQ1OT1 expression was remarkably upregulated in ESCC tissues and cell lines. Overexpression of KCNQ1OT1 markedly promoted ESCC cell proliferation, migration, and invasion and enhanced the expression of N-cadherin, MMP-2, and MMP-9, but inhibited apoptosis and E-cadherin expression in ESCC cell lines; KCNQ1OT1 knockdown exerted the opposite effects. KCNQ1OT1 could directly bind to miR-133b and suppress its expression, and miR-133b reversed the effects of KCNQ1OT1 overexpression in ESCC cells. MiR-133b reduced the expression of epidermal growth factor receptor (EGFR); further, KCNQ1OT1 activated the phosphatidylinositol 3-kinase/AKT serine/threonine kinase 1 (PI3K/AKT) signaling pathway by repressing miR-133b repression and indirectly upregulating EGFR. KCNQ1OT1 expression was positively correlated with EGFR mRNA expression and negatively correlated with miR-133b expression. CONCLUSION: KCNQ1OT1 facilitates ESCC progression by sponging miR-133b and activating the EGFR/PI3K/AKT pathway.

Expression of lncrna KCNQ1OT1 in DLBCL patients and its clinical significance / 肿瘤

Objective: To investigate the expression of long non-coding RNA (LncRNA) KCNQ1 overlapping transcript 1 (KCNQ1OT1) in the peripheral blood of patients with diffuse large B-cell lymphoma (DLBCL), and to analyze its clinical significance. Methods: The expression of LncRNA KCNQ1OT1 in the serum samples of 120 DLBCL patients and 55 non-tumor patients (as the control group) was detected by real-time fluorescent quantitative PCR. The correlations of LncRNA KCNQ1OT1 expression with the clinicopathological features and prognosis of DLBCL patients were analyzed. Results: The expression level of LncRNA KCNQ1OT1 in the serum of DLBCL patients was higher than that in the control group (P < 0.001). The expression of LncRNA KCNQ1OT1 was correlated with the tumor size, Ann Arbor stage, B symptoms, chemosensitivity and the international prognostic index (IPI) (all P < 0.001). The progression-free survival (PFS) time and the overall survival (OS) time of DLBCL patients with LncRNA KCNQ1OT1 high expression were significantly shorter than those with LncRNA KCNQ1OT1 low expression (both P < 0.001). Ann Arbor stage, LncRNA KCNQ1OT1 expression, IPI and chemosensitivity were the independent prognosis factors for DLBCL patients (all P < 0.01). Conclusion: LncRNA KCNQ1OT1 is highly expressed in the serum of patients with DLBCL, and is one of independent prognosis factors for DLBCL patients.