d histidine powders  (Chem Impex International)

Journal: Journal of Cellular and Molecular Medicine

Article Title: Long intergenic non‐coding RNA Linc00485 promotes lung cancer progression by modulating miR‐298/c‐Myc axis

doi: 10.1111/jcmm.16036

Figure Lengend Snippet: Primers for real‐time PCR

Article Snippet: Wild‐type (WT) Linc00485 (WT‐00485), mutant Linc00485 (MUT‐00485), WT c‐Myc 3′‐UTR (WT‐c‐Myc), and mutant c‐Myc 3′‐UTR (MUT‐c‐Myc) plasmid vectors were constructed by Sangon Biotech (Shanghai) Co., Ltd.

Techniques: Sequencing

Journal: Journal of Cellular and Molecular Medicine

Article Title: Long intergenic non‐coding RNA Linc00485 promotes lung cancer progression by modulating miR‐298/c‐Myc axis

doi: 10.1111/jcmm.16036

Figure Lengend Snippet: Linc00485 is up‐regulated in lung cancer tissues and associated with poor prognosis in patients with lung cancer. Expression levels of Linc00485 in (A) paracancerous tissues and tumour tissues of patients with lung cancer, (B) tumour tissues of patients with lung cancer at different TNM stages, (C) tumour tissues of patients with metastatic lung cancer, and (D) tumour tissues of patients with relapsed lung cancer. E, Association of Linc00485 with 5‐y survival of patients with lung cancer. F, Linc00485 levels in human bronchial epithelial cell line BEAS‐2B and different types of lung cancer cells. G‐I, Intracellular sub‐localization of Linc00485 in various lung cancer cell lines. Data are expressed as mean ± SD (n = 3). Differences between two groups were analysed by Student's t test. Comparisons among multiple groups with uniform variance were made using ANOVA with post hoc least significant difference test. L, lymphatic metastasis; N, adjacent normal tissues; R, relapse; T, tumour tissues. * P < 0.05, ** P < 0.01, *** P < 0.001 compared with the control group

Article Snippet: Wild‐type (WT) Linc00485 (WT‐00485), mutant Linc00485 (MUT‐00485), WT c‐Myc 3′‐UTR (WT‐c‐Myc), and mutant c‐Myc 3′‐UTR (MUT‐c‐Myc) plasmid vectors were constructed by Sangon Biotech (Shanghai) Co., Ltd.

Techniques: Expressing

Journal: Journal of Cellular and Molecular Medicine

Article Title: Long intergenic non‐coding RNA Linc00485 promotes lung cancer progression by modulating miR‐298/c‐Myc axis

doi: 10.1111/jcmm.16036

Figure Lengend Snippet: Linc00485 silencing significantly inhibited and Linc00485 overexpression promoted lung cancer cell proliferation. A, Efficiency of Linc00485 silencing as detected by quantitative polymerase chain reaction (qPCR) in A549 cells. B, Cell viability measured by Cell counting kit‐8 (CCK‐8) assay in A549 cells with Linc00485 silencing. C, Colony formation assay; the numbers of A549 cells in each colony were counted (n = 10 fields of view). D, Ki‐67‐positive rate of A549 cells with Linc00485 silencing, as detected by flow cytometry. E, Efficiency of Linc00485 overexpression, as detected by qPCR in A549 cells. F, Overexpression of Linc00485 promoted the viability of A549 cells. G, Cell number in each colony was significantly increased in A549 cells with Linc00485 overexpression. H, Linc00485 overexpression in A549 cells elevated the Ki‐67‐positive rate. I, Efficiency of Linc00485 silencing, as detected by qPCR in H460 cells. J, Linc00485 silencing significantly inhibited the viability of H460 cells. K, Efficiency of Linc00485 silencing, as detected by qPCR in H1975 cells. L, Linc00485 silencing markedly attenuated the viability of H1975 cells. M, Tumour volume was monitored weekly for 5 wk. N, Tumour weight was measured 35 d after implantation. O, Expression levels of cell proliferation‐related genes (Bax, Bid, P53, Bcl‐2, CDK4, and CDK6) in xenograft tumour tissues. Data are from three independent experiments and were analysed using Student's t test. Error bars represent SD. OE‐00485, overexpression of LincRNA 00485; sh‐00485, lentivirus‐mediated short hairpin RNA knockdown of LincRNA 00485; si‐00485, small interfering RNA for LincRNA 00485. * P < 0.05, ** P < 0.01, *** P < 0.001 compared with the control group

Article Snippet: Wild‐type (WT) Linc00485 (WT‐00485), mutant Linc00485 (MUT‐00485), WT c‐Myc 3′‐UTR (WT‐c‐Myc), and mutant c‐Myc 3′‐UTR (MUT‐c‐Myc) plasmid vectors were constructed by Sangon Biotech (Shanghai) Co., Ltd.

Techniques: Over Expression, Real-time Polymerase Chain Reaction, Cell Counting, CCK-8 Assay, Colony Assay, Flow Cytometry, Expressing, shRNA, Small Interfering RNA

Journal: Journal of Cellular and Molecular Medicine

Article Title: Long intergenic non‐coding RNA Linc00485 promotes lung cancer progression by modulating miR‐298/c‐Myc axis

doi: 10.1111/jcmm.16036

Figure Lengend Snippet: Silencing of Linc00485 suppresses and overexpression of Linc00485 facilitates migration and invasion of lung cancer cells. A‐F, Linc00485 silencing repressed the migration and invasion of (A, B) A549 cells, (C, D) H460 cells, and (E, F) H1975 cells. G and H, Linc00485 overexpression promoted the migratory and invasive abilities of A549 cells. I and J, Linc00485 levels and mRNA levels of E‐cadherin, CK‐19, N‐cadherin, vimentin, MMP‐2, and MMP‐9 in Linc00485‐silenced A549 cells and mouse tumour tissues (n = 6 mice in each group). Data are from three independent experiments and were analysed using Student's t test. Error bars represent SD. Scale bar = 50 μm. sh‐00485, lentivirus‐mediated short hairpin RNA knockdown of LincRNA 00485. * P < 0.05, ** P < 0.01, *** P < 0.001 compared with the control group

Article Snippet: Wild‐type (WT) Linc00485 (WT‐00485), mutant Linc00485 (MUT‐00485), WT c‐Myc 3′‐UTR (WT‐c‐Myc), and mutant c‐Myc 3′‐UTR (MUT‐c‐Myc) plasmid vectors were constructed by Sangon Biotech (Shanghai) Co., Ltd.

Techniques: Over Expression, Migration, shRNA

Journal: Journal of Cellular and Molecular Medicine

Article Title: Long intergenic non‐coding RNA Linc00485 promotes lung cancer progression by modulating miR‐298/c‐Myc axis

doi: 10.1111/jcmm.16036

Figure Lengend Snippet: Linc00485 directly binds to miR‐298. A, Schematic of the binding sites between Linc00485 and miR‐298. B‐E, Expression levels of miR‐298 (B) in paracancerous tissues and tumour tissues of patients with lung cancer, (C) in patients with lung cancer at different TNM stages, (D) in patients with relapsed lung cancer, and (E) in patients with metastatic lung cancer. F, Lung cancer patients with low expression of miR‐298 had significantly better outcomes. G, Pearson correlation analysis showed that miR‐298 expression had a reverse correlation with Linc00485 expression in adjacent normal tissues (left) and tumour tissues (right) of patients with lung cancer, respectively. H, Expression levels of miR‐298 in lung epithelial cells and lung cancer cells. I and J, miR‐298 expression after (I) overexpression or (J) silencing of Linc00485 in A549 cells. K, RNA immunoprecipitation assay confirmed the interaction relationship between Linc00485 and miR‐298 in A549 cells. L, Dual‐luciferase reporter assay revealed that wild‐type (WT) Linc00485 (left) could bind to miR‐298, but mutant Linc00485 (right) could not. M, Expression of miR‐298 was significantly up‐regulated in xenograft tumour tissues with Linc00485 silencing (n = 6 mice in each group). Data are from three independent experiments and were analysed using Student's t test or ANOVA with post hoc least significant difference test. Correlation analysis was used to analyse the 5‐y survival of patients with lung cancer. Error bars represent SD. OE‐00485, overexpression of LincRNA 00485; sh‐00485, lentivirus‐mediated short hairpin RNA knockdown of LincRNA 00485. L, lymphatic metastasis; N, adjacent normal tissues; R, relapse; T, tumour tissues. * P < 0.05, ** P < 0.01, *** P < 0.001 compared with the control group

Article Snippet: Wild‐type (WT) Linc00485 (WT‐00485), mutant Linc00485 (MUT‐00485), WT c‐Myc 3′‐UTR (WT‐c‐Myc), and mutant c‐Myc 3′‐UTR (MUT‐c‐Myc) plasmid vectors were constructed by Sangon Biotech (Shanghai) Co., Ltd.

Techniques: Binding Assay, Expressing, Over Expression, Immunoprecipitation, Luciferase, Reporter Assay, Mutagenesis, shRNA

Journal: Journal of Cellular and Molecular Medicine

Article Title: Long intergenic non‐coding RNA Linc00485 promotes lung cancer progression by modulating miR‐298/c‐Myc axis

doi: 10.1111/jcmm.16036

Figure Lengend Snippet: Linc00485/miR‐298 axis modulates proliferation, migration and invasion processes of lung cancer cells. A, Transfection efficiency of miR‐298 mimic or inhibitor in A549 cells. (B, C) Proliferation of A549 cells transfected with miR‐298 mimic or inhibitor measured by (B) Cell counting kit‐8 (CCK‐8) assay and (C) Ki‐67 staining. D‐F, The migratory and invasive capabilities of A549 cells transfected with miR‐298 mimic or inhibitor were evaluated by (D, E) an in vitro scratch assay, (F) transwell migration and invasion assays. G, CCK‐8 assay, (H) colony formation assays, and (I) Ki‐67 staining indicated that Linc00485 knockdown in combination with miR‐298 mimic transfection synergistically inhibited lung cancer cell viability, whereas miR‐298 inhibitor treatment could partially reverse the results of Linc00485 silencing. J and K, Cell motility and invasiveness were reduced in A549 cells after co‐transfection with sh‐00485 and miR‐298 mimic, whereas miR‐298 inhibitor reversed Linc00485‐knockdown‐induced changes compared with A549 cells treated with sh‐00485 alone. Data are from three independent experiments and were analysed using Student's t test or ANOVA with post hoc LSD test. Error bars represent SD. Scale bar = 50 μm. sh‐00485, lentivirus‐mediated short hairpin RNA knockdown of LincRNA 00485. * P < 0.05, ** P < 0.01, *** P < 0.001 compared with the control group

Article Snippet: Wild‐type (WT) Linc00485 (WT‐00485), mutant Linc00485 (MUT‐00485), WT c‐Myc 3′‐UTR (WT‐c‐Myc), and mutant c‐Myc 3′‐UTR (MUT‐c‐Myc) plasmid vectors were constructed by Sangon Biotech (Shanghai) Co., Ltd.

Techniques: Migration, Transfection, Cell Counting, CCK-8 Assay, Staining, In Vitro, Wound Healing Assay, Cotransfection, shRNA

Journal: Journal of Cellular and Molecular Medicine

Article Title: Long intergenic non‐coding RNA Linc00485 promotes lung cancer progression by modulating miR‐298/c‐Myc axis

doi: 10.1111/jcmm.16036

Figure Lengend Snippet: The c‐Myc gene is the downstream target of the Linc00485/miR‐298 axis. A, Schematic of the binding sites between miR‐298 and c‐Myc. B, Expression of c‐Myc in tumour tissues and paired normal tissues from patients with lung cancer, and its relationship with (C) TNM stage, (D) recurrence, (E) lymphatic metastasis and (F) 5‐y survival rate. G and H, Pearson correlation analysis indicated that (G) c‐Myc expression was positively correlated with the expression level of Linc00485 but (H) negatively correlated with miR‐298 levels in adjacent normal tissue (left) and tumour tissue (right). I, Expression levels of c‐Myc in lung epithelial cells and lung cancer cells. J, c‐Myc gene expression was dramatically down‐regulated in Linc00485‐silenced in A549 cells but (K) markedly up‐regulated in Linc00485‐overexpressing A549 cells compared with control cells. L, Interaction between c‐Myc mRNA (left) and miR‐298 (right) in A549 cells detected by RNA immunoprecipitation (RIP) assay. (M) Dual‐fluorescein reporter experiments confirmed that the wild‐type (WT) c‐Myc 3′‐untranslated region (UTR) (left) bound to miR‐298, but the mutant c‐Myc 3′‐UTR (right) could not. (N) Expression of c‐Myc in xenograft tumour tissues was significantly reduced after silencing of Linc00485 (n = 6 mice in each group). Data are from three independent experiments and were analysed using Student's t test or ANOVA with post hoc LSD test. Correlation analysis was used to analyse the 5‐y survival of patients with lung cancer. Error bars represent SD. L, lymphatic metastasis; N, adjacent normal tissues; R, relapse; T, tumour tissues. * P < 0.05, ** P < 0.01, *** P < 0.001 compared with the control group

Article Snippet: Wild‐type (WT) Linc00485 (WT‐00485), mutant Linc00485 (MUT‐00485), WT c‐Myc 3′‐UTR (WT‐c‐Myc), and mutant c‐Myc 3′‐UTR (MUT‐c‐Myc) plasmid vectors were constructed by Sangon Biotech (Shanghai) Co., Ltd.

Techniques: Binding Assay, Expressing, Immunoprecipitation, Mutagenesis

Journal: Journal of Cellular and Molecular Medicine

Article Title: Long intergenic non‐coding RNA Linc00485 promotes lung cancer progression by modulating miR‐298/c‐Myc axis

doi: 10.1111/jcmm.16036

Figure Lengend Snippet: Linc00485/miR‐298/c‐Myc axis exerts pro‐tumour effects on proliferation, migration, and invasion of lung cancer cells. A‐C, Cell proliferation ability of A549 cells co‐transfected with sh‐00485 and si‐c‐Myc or OE‐c‐Myc, as tested by (A) CCK‐8, (B) colony formation assay, and (C) Ki‐67 staining. D‐F, Migratory and invasive ability of A549 cells co‐transfected with sh‐00485 and si‐c‐Myc or OE‐c‐Myc, measured using (D) an in vitro scratch assay, and (E, F) transwell migration and invasion assays. G‐I, Viability, (J‐L) migration, and invasion of A549 cells co‐transfected with miR‐298 mimic and si‐c‐Myc or OE‐c‐Myc. M‐P, Effects of Linc00485 silencing and c‐Myc inhibitor 10074‐G5 treatment (3 μmol/L, 24 h) or overexpression of c‐Myc on the proliferation, migration, and invasion of A549 cells. Q‐T, Effects of Linc00485 overexpression in combination with 10074‐G5 treatment (3 μmol/L, 24 h) on the proliferation, migration, and invasion of A549 cells. U, Schematic diagram illustrating the role of Linc00485 in lung cancer. Data are from three independent experiments and were analysed using ANOVA with post hoc LSD test. Error bars represent SD. Scale bar = 50 μm. OE‐00485, overexpression of LincRNA 00485; OE‐c‐Myc, overexpression of c‐Myc; sh‐00485, lentivirus‐mediated short hairpin RNA knockdown of LincRNA 00485; si‐c‐Myc, small interfering RNA for c‐Myc. * P < 0.05, ** P < 0.01 compared with the control group

Article Snippet: Wild‐type (WT) Linc00485 (WT‐00485), mutant Linc00485 (MUT‐00485), WT c‐Myc 3′‐UTR (WT‐c‐Myc), and mutant c‐Myc 3′‐UTR (MUT‐c‐Myc) plasmid vectors were constructed by Sangon Biotech (Shanghai) Co., Ltd.

Techniques: Migration, Transfection, CCK-8 Assay, Colony Assay, Staining, In Vitro, Wound Healing Assay, Over Expression, shRNA, Small Interfering RNA

Journal: Journal of Cellular and Molecular Medicine

Article Title: Long intergenic non‐coding RNA Linc00485 promotes lung cancer progression by modulating miR‐298/c‐Myc axis

doi: 10.1111/jcmm.16036

Figure Lengend Snippet: Primers for real‐time PCR

Article Snippet: Wild‐type (WT) Linc00485 (WT‐00485), mutant Linc00485 (MUT‐00485), WT c‐Myc 3′‐UTR (WT‐c‐Myc), and mutant c‐Myc 3′‐UTR (MUT‐c‐Myc) plasmid vectors were constructed by Sangon Biotech (Shanghai) Co., Ltd.

Techniques: Sequencing

Journal: Journal of Cellular and Molecular Medicine

Article Title: Long intergenic non‐coding RNA Linc00485 promotes lung cancer progression by modulating miR‐298/c‐Myc axis

doi: 10.1111/jcmm.16036

Figure Lengend Snippet: Linc00485 is up‐regulated in lung cancer tissues and associated with poor prognosis in patients with lung cancer. Expression levels of Linc00485 in (A) paracancerous tissues and tumour tissues of patients with lung cancer, (B) tumour tissues of patients with lung cancer at different TNM stages, (C) tumour tissues of patients with metastatic lung cancer, and (D) tumour tissues of patients with relapsed lung cancer. E, Association of Linc00485 with 5‐y survival of patients with lung cancer. F, Linc00485 levels in human bronchial epithelial cell line BEAS‐2B and different types of lung cancer cells. G‐I, Intracellular sub‐localization of Linc00485 in various lung cancer cell lines. Data are expressed as mean ± SD (n = 3). Differences between two groups were analysed by Student's t test. Comparisons among multiple groups with uniform variance were made using ANOVA with post hoc least significant difference test. L, lymphatic metastasis; N, adjacent normal tissues; R, relapse; T, tumour tissues. * P < 0.05, ** P < 0.01, *** P < 0.001 compared with the control group

Article Snippet: Wild‐type (WT) Linc00485 (WT‐00485), mutant Linc00485 (MUT‐00485), WT c‐Myc 3′‐UTR (WT‐c‐Myc), and mutant c‐Myc 3′‐UTR (MUT‐c‐Myc) plasmid vectors were constructed by Sangon Biotech (Shanghai) Co., Ltd.

Techniques: Expressing

Journal: Journal of Cellular and Molecular Medicine

Article Title: Long intergenic non‐coding RNA Linc00485 promotes lung cancer progression by modulating miR‐298/c‐Myc axis

doi: 10.1111/jcmm.16036

Figure Lengend Snippet: Linc00485 silencing significantly inhibited and Linc00485 overexpression promoted lung cancer cell proliferation. A, Efficiency of Linc00485 silencing as detected by quantitative polymerase chain reaction (qPCR) in A549 cells. B, Cell viability measured by Cell counting kit‐8 (CCK‐8) assay in A549 cells with Linc00485 silencing. C, Colony formation assay; the numbers of A549 cells in each colony were counted (n = 10 fields of view). D, Ki‐67‐positive rate of A549 cells with Linc00485 silencing, as detected by flow cytometry. E, Efficiency of Linc00485 overexpression, as detected by qPCR in A549 cells. F, Overexpression of Linc00485 promoted the viability of A549 cells. G, Cell number in each colony was significantly increased in A549 cells with Linc00485 overexpression. H, Linc00485 overexpression in A549 cells elevated the Ki‐67‐positive rate. I, Efficiency of Linc00485 silencing, as detected by qPCR in H460 cells. J, Linc00485 silencing significantly inhibited the viability of H460 cells. K, Efficiency of Linc00485 silencing, as detected by qPCR in H1975 cells. L, Linc00485 silencing markedly attenuated the viability of H1975 cells. M, Tumour volume was monitored weekly for 5 wk. N, Tumour weight was measured 35 d after implantation. O, Expression levels of cell proliferation‐related genes (Bax, Bid, P53, Bcl‐2, CDK4, and CDK6) in xenograft tumour tissues. Data are from three independent experiments and were analysed using Student's t test. Error bars represent SD. OE‐00485, overexpression of LincRNA 00485; sh‐00485, lentivirus‐mediated short hairpin RNA knockdown of LincRNA 00485; si‐00485, small interfering RNA for LincRNA 00485. * P < 0.05, ** P < 0.01, *** P < 0.001 compared with the control group

Article Snippet: Wild‐type (WT) Linc00485 (WT‐00485), mutant Linc00485 (MUT‐00485), WT c‐Myc 3′‐UTR (WT‐c‐Myc), and mutant c‐Myc 3′‐UTR (MUT‐c‐Myc) plasmid vectors were constructed by Sangon Biotech (Shanghai) Co., Ltd.

Techniques: Over Expression, Real-time Polymerase Chain Reaction, Cell Counting, CCK-8 Assay, Colony Assay, Flow Cytometry, Expressing, shRNA, Small Interfering RNA

Journal: Journal of Cellular and Molecular Medicine

Article Title: Long intergenic non‐coding RNA Linc00485 promotes lung cancer progression by modulating miR‐298/c‐Myc axis

doi: 10.1111/jcmm.16036

Figure Lengend Snippet: Silencing of Linc00485 suppresses and overexpression of Linc00485 facilitates migration and invasion of lung cancer cells. A‐F, Linc00485 silencing repressed the migration and invasion of (A, B) A549 cells, (C, D) H460 cells, and (E, F) H1975 cells. G and H, Linc00485 overexpression promoted the migratory and invasive abilities of A549 cells. I and J, Linc00485 levels and mRNA levels of E‐cadherin, CK‐19, N‐cadherin, vimentin, MMP‐2, and MMP‐9 in Linc00485‐silenced A549 cells and mouse tumour tissues (n = 6 mice in each group). Data are from three independent experiments and were analysed using Student's t test. Error bars represent SD. Scale bar = 50 μm. sh‐00485, lentivirus‐mediated short hairpin RNA knockdown of LincRNA 00485. * P < 0.05, ** P < 0.01, *** P < 0.001 compared with the control group

Article Snippet: Wild‐type (WT) Linc00485 (WT‐00485), mutant Linc00485 (MUT‐00485), WT c‐Myc 3′‐UTR (WT‐c‐Myc), and mutant c‐Myc 3′‐UTR (MUT‐c‐Myc) plasmid vectors were constructed by Sangon Biotech (Shanghai) Co., Ltd.

Techniques: Over Expression, Migration, shRNA

Journal: Journal of Cellular and Molecular Medicine

Article Title: Long intergenic non‐coding RNA Linc00485 promotes lung cancer progression by modulating miR‐298/c‐Myc axis

doi: 10.1111/jcmm.16036

Figure Lengend Snippet: Linc00485 directly binds to miR‐298. A, Schematic of the binding sites between Linc00485 and miR‐298. B‐E, Expression levels of miR‐298 (B) in paracancerous tissues and tumour tissues of patients with lung cancer, (C) in patients with lung cancer at different TNM stages, (D) in patients with relapsed lung cancer, and (E) in patients with metastatic lung cancer. F, Lung cancer patients with low expression of miR‐298 had significantly better outcomes. G, Pearson correlation analysis showed that miR‐298 expression had a reverse correlation with Linc00485 expression in adjacent normal tissues (left) and tumour tissues (right) of patients with lung cancer, respectively. H, Expression levels of miR‐298 in lung epithelial cells and lung cancer cells. I and J, miR‐298 expression after (I) overexpression or (J) silencing of Linc00485 in A549 cells. K, RNA immunoprecipitation assay confirmed the interaction relationship between Linc00485 and miR‐298 in A549 cells. L, Dual‐luciferase reporter assay revealed that wild‐type (WT) Linc00485 (left) could bind to miR‐298, but mutant Linc00485 (right) could not. M, Expression of miR‐298 was significantly up‐regulated in xenograft tumour tissues with Linc00485 silencing (n = 6 mice in each group). Data are from three independent experiments and were analysed using Student's t test or ANOVA with post hoc least significant difference test. Correlation analysis was used to analyse the 5‐y survival of patients with lung cancer. Error bars represent SD. OE‐00485, overexpression of LincRNA 00485; sh‐00485, lentivirus‐mediated short hairpin RNA knockdown of LincRNA 00485. L, lymphatic metastasis; N, adjacent normal tissues; R, relapse; T, tumour tissues. * P < 0.05, ** P < 0.01, *** P < 0.001 compared with the control group

Article Snippet: Wild‐type (WT) Linc00485 (WT‐00485), mutant Linc00485 (MUT‐00485), WT c‐Myc 3′‐UTR (WT‐c‐Myc), and mutant c‐Myc 3′‐UTR (MUT‐c‐Myc) plasmid vectors were constructed by Sangon Biotech (Shanghai) Co., Ltd.

Techniques: Binding Assay, Expressing, Over Expression, Immunoprecipitation, Luciferase, Reporter Assay, Mutagenesis, shRNA

Journal: Journal of Cellular and Molecular Medicine

Article Title: Long intergenic non‐coding RNA Linc00485 promotes lung cancer progression by modulating miR‐298/c‐Myc axis

doi: 10.1111/jcmm.16036

Figure Lengend Snippet: Linc00485/miR‐298 axis modulates proliferation, migration and invasion processes of lung cancer cells. A, Transfection efficiency of miR‐298 mimic or inhibitor in A549 cells. (B, C) Proliferation of A549 cells transfected with miR‐298 mimic or inhibitor measured by (B) Cell counting kit‐8 (CCK‐8) assay and (C) Ki‐67 staining. D‐F, The migratory and invasive capabilities of A549 cells transfected with miR‐298 mimic or inhibitor were evaluated by (D, E) an in vitro scratch assay, (F) transwell migration and invasion assays. G, CCK‐8 assay, (H) colony formation assays, and (I) Ki‐67 staining indicated that Linc00485 knockdown in combination with miR‐298 mimic transfection synergistically inhibited lung cancer cell viability, whereas miR‐298 inhibitor treatment could partially reverse the results of Linc00485 silencing. J and K, Cell motility and invasiveness were reduced in A549 cells after co‐transfection with sh‐00485 and miR‐298 mimic, whereas miR‐298 inhibitor reversed Linc00485‐knockdown‐induced changes compared with A549 cells treated with sh‐00485 alone. Data are from three independent experiments and were analysed using Student's t test or ANOVA with post hoc LSD test. Error bars represent SD. Scale bar = 50 μm. sh‐00485, lentivirus‐mediated short hairpin RNA knockdown of LincRNA 00485. * P < 0.05, ** P < 0.01, *** P < 0.001 compared with the control group

Article Snippet: Wild‐type (WT) Linc00485 (WT‐00485), mutant Linc00485 (MUT‐00485), WT c‐Myc 3′‐UTR (WT‐c‐Myc), and mutant c‐Myc 3′‐UTR (MUT‐c‐Myc) plasmid vectors were constructed by Sangon Biotech (Shanghai) Co., Ltd.

Techniques: Migration, Transfection, Cell Counting, CCK-8 Assay, Staining, In Vitro, Wound Healing Assay, Cotransfection, shRNA

Journal: Journal of Cellular and Molecular Medicine

Article Title: Long intergenic non‐coding RNA Linc00485 promotes lung cancer progression by modulating miR‐298/c‐Myc axis

doi: 10.1111/jcmm.16036

Figure Lengend Snippet: The c‐Myc gene is the downstream target of the Linc00485/miR‐298 axis. A, Schematic of the binding sites between miR‐298 and c‐Myc. B, Expression of c‐Myc in tumour tissues and paired normal tissues from patients with lung cancer, and its relationship with (C) TNM stage, (D) recurrence, (E) lymphatic metastasis and (F) 5‐y survival rate. G and H, Pearson correlation analysis indicated that (G) c‐Myc expression was positively correlated with the expression level of Linc00485 but (H) negatively correlated with miR‐298 levels in adjacent normal tissue (left) and tumour tissue (right). I, Expression levels of c‐Myc in lung epithelial cells and lung cancer cells. J, c‐Myc gene expression was dramatically down‐regulated in Linc00485‐silenced in A549 cells but (K) markedly up‐regulated in Linc00485‐overexpressing A549 cells compared with control cells. L, Interaction between c‐Myc mRNA (left) and miR‐298 (right) in A549 cells detected by RNA immunoprecipitation (RIP) assay. (M) Dual‐fluorescein reporter experiments confirmed that the wild‐type (WT) c‐Myc 3′‐untranslated region (UTR) (left) bound to miR‐298, but the mutant c‐Myc 3′‐UTR (right) could not. (N) Expression of c‐Myc in xenograft tumour tissues was significantly reduced after silencing of Linc00485 (n = 6 mice in each group). Data are from three independent experiments and were analysed using Student's t test or ANOVA with post hoc LSD test. Correlation analysis was used to analyse the 5‐y survival of patients with lung cancer. Error bars represent SD. L, lymphatic metastasis; N, adjacent normal tissues; R, relapse; T, tumour tissues. * P < 0.05, ** P < 0.01, *** P < 0.001 compared with the control group

Article Snippet: Wild‐type (WT) Linc00485 (WT‐00485), mutant Linc00485 (MUT‐00485), WT c‐Myc 3′‐UTR (WT‐c‐Myc), and mutant c‐Myc 3′‐UTR (MUT‐c‐Myc) plasmid vectors were constructed by Sangon Biotech (Shanghai) Co., Ltd.

Techniques: Binding Assay, Expressing, Immunoprecipitation, Mutagenesis

Journal: Journal of Cellular and Molecular Medicine

Article Title: Long intergenic non‐coding RNA Linc00485 promotes lung cancer progression by modulating miR‐298/c‐Myc axis

doi: 10.1111/jcmm.16036

Figure Lengend Snippet: Linc00485/miR‐298/c‐Myc axis exerts pro‐tumour effects on proliferation, migration, and invasion of lung cancer cells. A‐C, Cell proliferation ability of A549 cells co‐transfected with sh‐00485 and si‐c‐Myc or OE‐c‐Myc, as tested by (A) CCK‐8, (B) colony formation assay, and (C) Ki‐67 staining. D‐F, Migratory and invasive ability of A549 cells co‐transfected with sh‐00485 and si‐c‐Myc or OE‐c‐Myc, measured using (D) an in vitro scratch assay, and (E, F) transwell migration and invasion assays. G‐I, Viability, (J‐L) migration, and invasion of A549 cells co‐transfected with miR‐298 mimic and si‐c‐Myc or OE‐c‐Myc. M‐P, Effects of Linc00485 silencing and c‐Myc inhibitor 10074‐G5 treatment (3 μmol/L, 24 h) or overexpression of c‐Myc on the proliferation, migration, and invasion of A549 cells. Q‐T, Effects of Linc00485 overexpression in combination with 10074‐G5 treatment (3 μmol/L, 24 h) on the proliferation, migration, and invasion of A549 cells. U, Schematic diagram illustrating the role of Linc00485 in lung cancer. Data are from three independent experiments and were analysed using ANOVA with post hoc LSD test. Error bars represent SD. Scale bar = 50 μm. OE‐00485, overexpression of LincRNA 00485; OE‐c‐Myc, overexpression of c‐Myc; sh‐00485, lentivirus‐mediated short hairpin RNA knockdown of LincRNA 00485; si‐c‐Myc, small interfering RNA for c‐Myc. * P < 0.05, ** P < 0.01 compared with the control group

Article Snippet: Wild‐type (WT) Linc00485 (WT‐00485), mutant Linc00485 (MUT‐00485), WT c‐Myc 3′‐UTR (WT‐c‐Myc), and mutant c‐Myc 3′‐UTR (MUT‐c‐Myc) plasmid vectors were constructed by Sangon Biotech (Shanghai) Co., Ltd.

Techniques: Migration, Transfection, CCK-8 Assay, Colony Assay, Staining, In Vitro, Wound Healing Assay, Over Expression, shRNA, Small Interfering RNA