mir 33a 5p mimics  (MedChemExpress)

Journal: Aging (Albany NY)

Article Title: miR-33a-5p inhibits the progression of esophageal cancer through the DKK1-mediated Wnt/β-catenin pathway

doi: 10.18632/aging.203430

Figure Lengend Snippet: Clinical value of miR-33a-5p and DKK1 in esophageal cancer. ( A ) Expression of miR-33a-5p in esophageal cancer tissues; ( B ) Serum miR-33a-5p expression in esophageal cancer and ROC curve; ( C ) Expression of DKK1 in esophageal cancer; ( D ) Serum DKK1 expression in esophageal cancer and ROC curve; ( E ) Serum miR-33a-5p expression was correlated with esophageal cancer; ( F ) Serum DKK1 expression was correlated with the esophageal cancer; ( G ) Correlation between miR-33a-5p and DKK1 in esophageal cancer tissues; ( H ) Correlation between miR-33a-5p and DKK1 in serum of esophageal cancer patients; ( I ) Relationship between serum miR-33a-5p expression and 5-year OS of patients; ( J ) Relationship between serum DKK1 expression and 5-year OS of patients. The inter-group comparison was conducted by the independent sample t -test, mean ± SD, b P < 0.01 for inter-group comparisons.

Article Snippet: Transfection was performed when the confluence reached 70%. miR-33a-5p-mimics, miR-335-3-inhibitor, miR-33a-5p negative control sequence (miR-NC), DKK1 RNA negative control (si-NC), targeted inhibition of DKK1 RNA (si-DKK1) and targeted overexpression of DKK1 RNA (sh- DKK1) were synthesized and obtained from ThermoFisher Scientific, Shanghai, China.

Techniques: Expressing, Comparison

Journal: Aging (Albany NY)

Article Title: miR-33a-5p inhibits the progression of esophageal cancer through the DKK1-mediated Wnt/β-catenin pathway

doi: 10.18632/aging.203430

Figure Lengend Snippet: Occurrence value of serum miR-33a-5p and DKK1 in EC.

Article Snippet: Transfection was performed when the confluence reached 70%. miR-33a-5p-mimics, miR-335-3-inhibitor, miR-33a-5p negative control sequence (miR-NC), DKK1 RNA negative control (si-NC), targeted inhibition of DKK1 RNA (si-DKK1) and targeted overexpression of DKK1 RNA (sh- DKK1) were synthesized and obtained from ThermoFisher Scientific, Shanghai, China.

Techniques:

Journal: Aging (Albany NY)

Article Title: miR-33a-5p inhibits the progression of esophageal cancer through the DKK1-mediated Wnt/β-catenin pathway

doi: 10.18632/aging.203430

Figure Lengend Snippet: Correlation of miR-33a-5p and DKK1 with clinical characteristics of patients with EC.

Article Snippet: Transfection was performed when the confluence reached 70%. miR-33a-5p-mimics, miR-335-3-inhibitor, miR-33a-5p negative control sequence (miR-NC), DKK1 RNA negative control (si-NC), targeted inhibition of DKK1 RNA (si-DKK1) and targeted overexpression of DKK1 RNA (sh- DKK1) were synthesized and obtained from ThermoFisher Scientific, Shanghai, China.

Techniques:

Journal: Aging (Albany NY)

Article Title: miR-33a-5p inhibits the progression of esophageal cancer through the DKK1-mediated Wnt/β-catenin pathway

doi: 10.18632/aging.203430

Figure Lengend Snippet: Effects of miR-129-5p on biological function of esophageal cancer cells. ( A ) qRT-PCR detection of miR-33a-5p expression in esophageal cancer cells by; ( B ) qRT-PCR detection of miR-33a-5p expression in TE-1, Eca-109 after transfection with miR-NC, miR-33a-5p-mimics, miR-33a-5p-inhibitor; ( C ) Cell proliferation of TE-1 and Eca-109 by CCK-8; ( D ) Migration number of TE-1 and Eca-109 cells by Transwell; ( E ) Invasion number of TE-1 and Eca-109 cells by Transwell; ( F ) Apoptosis rate of TE-1 and Eca-109 cells by flow cytometry and the apoptosis diagram. The inter-group comparison was conducted by the independent sample t -test, multiple time point data were analyzed by repeated measures analysis of variance (ANOVA), mean ± SD, b P < 0.01, c P < 0.05, d P < 0.01, f P < 0.01, and h P < 0.01 for inter-group comparisons.

Article Snippet: Transfection was performed when the confluence reached 70%. miR-33a-5p-mimics, miR-335-3-inhibitor, miR-33a-5p negative control sequence (miR-NC), DKK1 RNA negative control (si-NC), targeted inhibition of DKK1 RNA (si-DKK1) and targeted overexpression of DKK1 RNA (sh- DKK1) were synthesized and obtained from ThermoFisher Scientific, Shanghai, China.

Techniques: Quantitative RT-PCR, Expressing, Transfection, CCK-8 Assay, Migration, Flow Cytometry, Comparison

Journal: Aging (Albany NY)

Article Title: miR-33a-5p inhibits the progression of esophageal cancer through the DKK1-mediated Wnt/β-catenin pathway

doi: 10.18632/aging.203430

Figure Lengend Snippet: DKK1 is identified as the target of miR-33a-5p. ( A ) The predicted binding sites between n DKK1 and miR-33a-5p. ( B ) Interaction between DKK1 and miR-33a-5p by dual luciferase reporter assay; ( C ) Western blot of DKK1 protein expression in TE-1 and Eca-109 cells after transfection with miR-NC, miR-33a-5p-mimics and miR-33a-5p-inhibitor and the protein diagram. The inter-group comparison was conducted by the independent sample t -test, mean ± SD, b P < 0.01, c P <0.05, and d P < 0.01 for inter-group comparisons.

Article Snippet: Transfection was performed when the confluence reached 70%. miR-33a-5p-mimics, miR-335-3-inhibitor, miR-33a-5p negative control sequence (miR-NC), DKK1 RNA negative control (si-NC), targeted inhibition of DKK1 RNA (si-DKK1) and targeted overexpression of DKK1 RNA (sh- DKK1) were synthesized and obtained from ThermoFisher Scientific, Shanghai, China.

Techniques: Binding Assay, Luciferase, Reporter Assay, Western Blot, Expressing, Transfection, Comparison

Journal: Aging (Albany NY)

Article Title: miR-33a-5p inhibits the progression of esophageal cancer through the DKK1-mediated Wnt/β-catenin pathway

doi: 10.18632/aging.203430

Figure Lengend Snippet: miR-33a-5p targeted DKK1-mediated Wnt/β-catenin pathway to affect biological behavior of esophageal cancer cells. ( A ) Cell proliferation of TE-1, Eca-109 by CCK-8; ( B ) Number of migration of TE-1 and Eca-109 cells by Transwell; ( C ) Number of invasion of TE-1 and Eca-109 cells by Transwell; ( D ) Cell apoptosis rates of TE-1and Eca-109 cells by Flow cytometry and the apoptosis map; ( E ) Western blot of Cyclin D1, c-jun, CD44, and c-Met protein expression in TE-1 and Eca-109 cells and the protein bands. The inter-group comparison was conducted by the independent sample t -test, multiple time point data were analyzed by repeated measures analysis of variance (ANOVA), mean ± SD, b P < 0.01, c P < 0.05, and d P < 0.01 for inter-group comparisons.

Article Snippet: Transfection was performed when the confluence reached 70%. miR-33a-5p-mimics, miR-335-3-inhibitor, miR-33a-5p negative control sequence (miR-NC), DKK1 RNA negative control (si-NC), targeted inhibition of DKK1 RNA (si-DKK1) and targeted overexpression of DKK1 RNA (sh- DKK1) were synthesized and obtained from ThermoFisher Scientific, Shanghai, China.

Techniques: CCK-8 Assay, Migration, Flow Cytometry, Western Blot, Expressing, Comparison

Journal: Aging (Albany NY)

Article Title: miR-33a-5p inhibits the progression of esophageal cancer through the DKK1-mediated Wnt/β-catenin pathway

doi: 10.18632/aging.203430

Figure Lengend Snippet: miR-33a-5p inhibits the tumor growth of EC in vivo . ( A – F ) The effect of miR-33a-5p on tumor growth of EC cells in vivo was analyzed by nude mice tumorigenicity assay by injected with the TE-1 cells treated with control mimic or miR-33a-5p mimic. ( A ) Representative images of dissected tumors from nude mice were presented. ( B ) The average tumor volume was calculated and shown. ( C ) The average tumor weight was calculated and shown. ( D ) The expression levels of miR-33a-5p were measured by qPCR in the tumor tissues of the mice. ( E ) The protein expression of DKK1 was assessed by Western blot analysis in the tumor tissues of the mice. ( F ) Western blot of Cyclin D1, c-jun, CD44, and c-Met protein expression in TE-1 and Eca-109 cells and the protein bands. N = 5. The inter-group comparison was conducted by the independent sample t -test, multiple time point data were analyzed by repeated measures analysis of variance (ANOVA), mean ± SD, b P < 0.01.

Article Snippet: Transfection was performed when the confluence reached 70%. miR-33a-5p-mimics, miR-335-3-inhibitor, miR-33a-5p negative control sequence (miR-NC), DKK1 RNA negative control (si-NC), targeted inhibition of DKK1 RNA (si-DKK1) and targeted overexpression of DKK1 RNA (sh- DKK1) were synthesized and obtained from ThermoFisher Scientific, Shanghai, China.

Techniques: In Vivo, Tumorigenicity Assay, Injection, Control, Expressing, Western Blot, Comparison

Journal: International Journal of Molecular Sciences

Article Title: How miR-31-5p and miR-33a-5p Regulates SP1/CX43 Expression in Osteoarthritis Disease: Preliminary Insights

doi: 10.3390/ijms22052471

Figure Lengend Snippet: Investigation of osteoblasts (OB) and chondrocytes (CH) isolated by tissue debris of mild OA and severe OA patients through qRT-PCR expression analysis of ( A ) miR-31-5p and miR-33a-5p; ( B ) miR-31-5p OB targets genes: RHO and HIF-1AN; and ( C ) miR-33a-5p CH targets genes: COL2A1 and HMGA2. Quantitative RT-PCR data are expressed as fold of change (FOI) in miRNAs or gene expression (2 −ΔΔCt ) occurred in severe OA-derived cells vs mild OA-derived cells. Data reported were analysed by Student t test: *, p < 0.05, **, p < 0.005, ***, p < 0.0005 between experimental group and represented as mean ± SD ( n = 4).

Article Snippet: Briefly, cells seeded at 150,000 cells/cm 2 were transfected with 15 pmoles/mL of: has-mir-31-5p mimic (4464066-MC11465, mirVana miRNA mimics Life Technologies Italia, Monza, Italy); has-mir-31-3p mimic (4464066-MC12887, mirVana miRNA mimics-Life Technologies, Monza, Italy); has-miR-31-5p inhibitor (4464084-MH11465, mirVana miRNA inhibitors-Life technologies Italia, Monza, Italy); hsa-mir-33a-5p mimic (4464066-MC12410, mirVana miRNA mimics-Life Technologies, Monza, Italy); hsa-mir-33a-3p mimic (4464066-MC12607, mirVana miRNA mimicsLife Technologies, Monza, Italy); hsa-mir-33a-5p inhibitor (4464084-MH12410, mirVana miRNA inhibitors -Life Technologies, Monza, Italy); and scrambled negative controls (4464058, mirVana negative control Life Technologies, Monza, Italy) for 24h.

Techniques: Isolation, Quantitative RT-PCR, Expressing, Derivative Assay

Journal: International Journal of Molecular Sciences

Article Title: How miR-31-5p and miR-33a-5p Regulates SP1/CX43 Expression in Osteoarthritis Disease: Preliminary Insights

doi: 10.3390/ijms22052471

Figure Lengend Snippet: Study of SP1 gene expression modulation by gain and loss of function assays. Osteoblasts (OB) and chondrocytes (CH) isolated by mild OA patients (mild OA) after 24 h of transfection with: miR-31-5p mimic and miR-31-3p mimic for OB cells, with miR-33a-5p mimic and miR-33a-3p mimic for CH cells and relatives scrambles. Cells were analyzed for the gene expression modulation of SP1 by qRT-PCR analysis. ( A ) The efficiency of miR-31-5p mimic and miR-31-3p mimic and relatives scrambles transfection were evaluated in transfected mild OA-derived OB versus untransfected ones. ( B ) The efficiency of miR-33a-5p mimic and miR-33a-3p and relatives scrambles transfection were evaluated in transfected mild OA-derived CH versus untransfected ones. SP1 gene expression evaluated in ( C ) OB isolated from mild OA patients transfected with mimic-31-5p and negative scramble, in ( D ) CH isolated from mild OA patients transfected with mimic-33a-5p and negative scramble. Quantitative RT-PCR data are expressed as fold of change (FOI) in miRNAs or gene expression (2 −ΔΔCt ) occurred in transfected mild OA isolated cells vs untransfected ones. Data reported were analysed by Student t test: *, p < 0.05, **, p < 0.005, ***, p < 0.0005 between experimental group and represented as mean ± SD ( n = 4).

Article Snippet: Briefly, cells seeded at 150,000 cells/cm 2 were transfected with 15 pmoles/mL of: has-mir-31-5p mimic (4464066-MC11465, mirVana miRNA mimics Life Technologies Italia, Monza, Italy); has-mir-31-3p mimic (4464066-MC12887, mirVana miRNA mimics-Life Technologies, Monza, Italy); has-miR-31-5p inhibitor (4464084-MH11465, mirVana miRNA inhibitors-Life technologies Italia, Monza, Italy); hsa-mir-33a-5p mimic (4464066-MC12410, mirVana miRNA mimics-Life Technologies, Monza, Italy); hsa-mir-33a-3p mimic (4464066-MC12607, mirVana miRNA mimicsLife Technologies, Monza, Italy); hsa-mir-33a-5p inhibitor (4464084-MH12410, mirVana miRNA inhibitors -Life Technologies, Monza, Italy); and scrambled negative controls (4464058, mirVana negative control Life Technologies, Monza, Italy) for 24h.

Techniques: Expressing, Isolation, Transfection, Quantitative RT-PCR, Derivative Assay

Journal: International Journal of Molecular Sciences

Article Title: How miR-31-5p and miR-33a-5p Regulates SP1/CX43 Expression in Osteoarthritis Disease: Preliminary Insights

doi: 10.3390/ijms22052471

Figure Lengend Snippet: Study of SP1 gene expression modulation by gain and loss of function assays. Osteoblasts (OB) and chondrocytes (CH) isolated by OA patients after 24 h of transfection with miR-31-5p inhibitor for OB cells and with miR-33a-5p inhibitor for CH cells and relatives scrambles were analyzed for the gene expression of SP1 by qRT-PCR analysis. ( A ) The efficiency of miR-31-5p inhibitor and relative scrambles transfection was evaluated in transfected severe OA-derived OB versus untransfected ones. ( B ) The efficiency of miR-33a-5p inhibitor and relatives scramble transfection was evaluated in transfected severe OA-derived CH versus untransfected ones. SP1 gene expression evaluated in ( C ) OB isolated from severe OA patients transfected with miR31-5p inhibitor and negative scramble, and in ( D ) CH isolated from severe OA patients transfected with miR-33a-5p inhibitor and negative scramble. Quantitative RT-PCR data are expressed as fold of change (FOI) in miRNAs or gene expression (2 −ΔΔCt ) occurred in transfected OA derived cells vs untransfected ones. Data reported were analysed by Student t test: *, p < 0.05, **, p < 0.005, between experimental group and represented as mean ± SD ( n = 4).

Article Snippet: Briefly, cells seeded at 150,000 cells/cm 2 were transfected with 15 pmoles/mL of: has-mir-31-5p mimic (4464066-MC11465, mirVana miRNA mimics Life Technologies Italia, Monza, Italy); has-mir-31-3p mimic (4464066-MC12887, mirVana miRNA mimics-Life Technologies, Monza, Italy); has-miR-31-5p inhibitor (4464084-MH11465, mirVana miRNA inhibitors-Life technologies Italia, Monza, Italy); hsa-mir-33a-5p mimic (4464066-MC12410, mirVana miRNA mimics-Life Technologies, Monza, Italy); hsa-mir-33a-3p mimic (4464066-MC12607, mirVana miRNA mimicsLife Technologies, Monza, Italy); hsa-mir-33a-5p inhibitor (4464084-MH12410, mirVana miRNA inhibitors -Life Technologies, Monza, Italy); and scrambled negative controls (4464058, mirVana negative control Life Technologies, Monza, Italy) for 24h.

Techniques: Expressing, Isolation, Transfection, Quantitative RT-PCR, Derivative Assay

Journal: International Journal of Molecular Sciences

Article Title: How miR-31-5p and miR-33a-5p Regulates SP1/CX43 Expression in Osteoarthritis Disease: Preliminary Insights

doi: 10.3390/ijms22052471

Figure Lengend Snippet: Our hypothesis of the molecular mechanism of miR-31-5p and miR-33a-5p on SP1/CX43 regulations in OA condition.

Article Snippet: Briefly, cells seeded at 150,000 cells/cm 2 were transfected with 15 pmoles/mL of: has-mir-31-5p mimic (4464066-MC11465, mirVana miRNA mimics Life Technologies Italia, Monza, Italy); has-mir-31-3p mimic (4464066-MC12887, mirVana miRNA mimics-Life Technologies, Monza, Italy); has-miR-31-5p inhibitor (4464084-MH11465, mirVana miRNA inhibitors-Life technologies Italia, Monza, Italy); hsa-mir-33a-5p mimic (4464066-MC12410, mirVana miRNA mimics-Life Technologies, Monza, Italy); hsa-mir-33a-3p mimic (4464066-MC12607, mirVana miRNA mimicsLife Technologies, Monza, Italy); hsa-mir-33a-5p inhibitor (4464084-MH12410, mirVana miRNA inhibitors -Life Technologies, Monza, Italy); and scrambled negative controls (4464058, mirVana negative control Life Technologies, Monza, Italy) for 24h.

Techniques: