Journal: Toxicology

Article Title: Ptaquiloside reduces NK cell activities by enhancing metallothionein expression, which is prevented by selenium.

doi: 10.1016/j.tox.2012.12.010

Figure Lengend Snippet: Fig. 4. Ptaquiloside treatment increases Mt2 expression in splenic NK cells that was inhibited by selenium co-treatment. Splenic NK cells were isolated from mice treated with ptaquiloside (5.3 mg/kg) and/or selenium (1.3 mg/kg) by daily gavage for 14 days. The differential gene expression detected using microarray analysis (5 mice/group) was

Article Snippet: Next, splenic cell suspensions were prepared from the other six untreated mice, and the non-adherent cells were incubated or not with the TrueORFTM vector containing M. musculus Mt2 cDNA (OriGene) as described above.

Techniques: Expressing, Isolation, Gene Expression, Microarray

Journal: Toxicology

Article Title: Ptaquiloside reduces NK cell activities by enhancing metallothionein expression, which is prevented by selenium.

doi: 10.1016/j.tox.2012.12.010

Figure Lengend Snippet: Fig. 7. Overexpression of Mt2 and its role in splenic NK cells. (A) Note the reduction in the free zinc level in the Mt2-transfected NK cells (*p = 0.0343, Student’s t-test). (B) The NK cytotoxicity is also reduced in the cells overexpressing Mt2 (*p = 0.0260, Mann–Whitney test). The data represent the mean and the error, n = 6.

Article Snippet: Next, splenic cell suspensions were prepared from the other six untreated mice, and the non-adherent cells were incubated or not with the TrueORFTM vector containing M. musculus Mt2 cDNA (OriGene) as described above.

Techniques: Over Expression, Transfection, MANN-WHITNEY

Journal: Toxicology

Article Title: Ptaquiloside reduces NK cell activities by enhancing metallothionein expression, which is prevented by selenium.

doi: 10.1016/j.tox.2012.12.010

Figure Lengend Snippet: Fig. 4. Ptaquiloside treatment increases Mt2 expression in splenic NK cells that was inhibited by selenium co-treatment. Splenic NK cells were isolated from mice treated with ptaquiloside (5.3 mg/kg) and/or selenium (1.3 mg/kg) by daily gavage for 14 days. The differential gene expression detected using microarray analysis (5 mice/group) was

Article Snippet: Briefly, 5 × 105 cells suspended in OptiMEM I (Invitrogen) were incubated with or without 0.2 g of TrueORFTM vector containing a Mus musculus Mt2 cDNA (OriGene) mixture with 0.5 l Lipofectamine (Invitrogen) per well at 37 ◦C in a humidified atmosphere at 5% CO2, following the manufacturer’s instructions.

Techniques: Expressing, Isolation, Gene Expression, Microarray

Journal: Toxicology

Article Title: Ptaquiloside reduces NK cell activities by enhancing metallothionein expression, which is prevented by selenium.

doi: 10.1016/j.tox.2012.12.010

Figure Lengend Snippet: Fig. 7. Overexpression of Mt2 and its role in splenic NK cells. (A) Note the reduction in the free zinc level in the Mt2-transfected NK cells (*p = 0.0343, Student’s t-test). (B) The NK cytotoxicity is also reduced in the cells overexpressing Mt2 (*p = 0.0260, Mann–Whitney test). The data represent the mean and the error, n = 6.

Article Snippet: Briefly, 5 × 105 cells suspended in OptiMEM I (Invitrogen) were incubated with or without 0.2 g of TrueORFTM vector containing a Mus musculus Mt2 cDNA (OriGene) mixture with 0.5 l Lipofectamine (Invitrogen) per well at 37 ◦C in a humidified atmosphere at 5% CO2, following the manufacturer’s instructions.

Techniques: Over Expression, Transfection, MANN-WHITNEY

Journal: Molecular Cancer

Article Title: Tumor cell invasion of collagen matrices requires coordinate lipid agonist-induced G-protein and membrane-type matrix metalloproteinase-1-dependent signaling

doi: 10.1186/1476-4598-5-69

Figure Lengend Snippet: HEK293 cells transfected with MT1-MMP cDNA invade 3D collagen gels in response to LPA . (A) Tumor cell lysates were prepared for Western blot analysis. Lysates were probed for MT1-MMP to assess protein expression in the four tumor cell lines. Lysates were probed for Actin as a loading control. (B) HEK293 cells were transfected with the pAdTrack-CMV plasmid as a control, or plasmids encoding MT1-MMP, MT2-MMP, or MT3-MMP cDNA 24 hours prior to placement in invasion assays. Cells were allowed to invade 2.0 mg/ml collagen gels in the presence or absence of 1 μM LPA. Data are expressed as mean numbers of invading cells per HPF (20×) (± S.D.) from a minimum of 20 fields. (C) Lysates from HEK293 cells transfected with cDNAs encoding the designated genes were prepared for Western blot analysis and probed for GFP, MT1-MMP, MT2-MMP, MT3-MMP, or Actin as a loading control. TRK = pAdTrack-CMV, MT1 = MT1-MMP, MT2 = MT2-MMP, MT3 = MT3-MMP.

Article Snippet: A plasmid encoding human MT2-MMP (TC118648) was purchased from Origene (Rockville, MD).

Techniques: Transfection, Western Blot, Expressing, Plasmid Preparation

Journal: International Journal of Molecular Sciences

Article Title: Metallothionein Family Proteins as Regulators of Zinc Ions Synergistically Enhance the Anticancer Effect of Cannabidiol in Human Colorectal Cancer Cells

doi: 10.3390/ijms242316621

Figure Lengend Snippet: qPCR analysis of metallothionein gene family during CBD treatment. ( A ) qRT-PCR was performed to examine expression level changes in metallothionein gene family by CBD treatment in two colorectal cancer cell lines. MT family genes were upregulated by CBD treatment in a time-dependent manner (* p ≤ 0.05; ** p ≤ 0.01). ( B ) Western blot analysis was performed for 20 μM CBD-treated colorectal cancer cells. Expression levels of MTs and MT2A were increased by CBD treatment in a time-dependent manner.

Article Snippet: The expression vectors of MT1G and MT2A were purchased from Origene (RC204741 and RC202748, Rockville, MD, USA).

Techniques: Quantitative RT-PCR, Expressing, Western Blot

Journal: International Journal of Molecular Sciences

Article Title: Metallothionein Family Proteins as Regulators of Zinc Ions Synergistically Enhance the Anticancer Effect of Cannabidiol in Human Colorectal Cancer Cells

doi: 10.3390/ijms242316621

Figure Lengend Snippet: Immunostaining analysis of metallothionein proteins after CBD treatment. Cells were treated with 20 μM CBD for 48 h, stained with MT antibody (red), MT2A antibody (green) and DAPI (blue). MT and MT2A levels were significantly increased in the cytoplasmic region after CBD treatment. (**, p ≤ 0.01). Scale bar, 10 µm.

Article Snippet: The expression vectors of MT1G and MT2A were purchased from Origene (RC204741 and RC202748, Rockville, MD, USA).

Techniques: Immunostaining, Staining

Journal: International Journal of Molecular Sciences

Article Title: Metallothionein Family Proteins as Regulators of Zinc Ions Synergistically Enhance the Anticancer Effect of Cannabidiol in Human Colorectal Cancer Cells

doi: 10.3390/ijms242316621

Figure Lengend Snippet: Synergistic effect of MT overexpression on anticancer effect of CBD. ( A ) SW480 cells were transfected with MT1G overexpression vector and treated with 20 μM CBD for 48 h. Apoptosis cells were analyzed using flow cytometry after staining with Annexin V/PI-FITC. ( B ) SW480 cells were transfected with MT2A overexpression vector and treated with 20 μM CBD for 48 h. Apoptosis cells were analyzed using flow cytometry after staining with Annexin V/PI-FITC. In both transient transfection experiments, the dead cell population (blue rectangles) was increased in the group with overexpression of MTs and CBD treatment. (** p ≤ 0.01).

Article Snippet: The expression vectors of MT1G and MT2A were purchased from Origene (RC204741 and RC202748, Rockville, MD, USA).

Techniques: Over Expression, Transfection, Plasmid Preparation, Flow Cytometry, Staining

Journal: International Journal of Molecular Sciences

Article Title: Metallothionein Family Proteins as Regulators of Zinc Ions Synergistically Enhance the Anticancer Effect of Cannabidiol in Human Colorectal Cancer Cells

doi: 10.3390/ijms242316621

Figure Lengend Snippet: Effect of combination treatment of CBD and zinc ions on anticancer activity. ( A ) Cells were treated with zinc ions for different time periods. MT and MT2A proteins were increased by zinc ion treatment in two colorectal cancer cell lines. ( B ) Cell viability was examined after combination treatment with sublethal concentration of CBD and zinc ions using MTT assay. Cell viability was significantly decreased by combination treatment with 5 μM CBD and zinc ions. ( C ) Flow cytometry analysis showed that the dead cell population was increased by combination treatment with 5 μM CBD and zinc ions. ( D ) Apoptosis cells were analyzed using flow cytometry after Annexin V/PI dual staining following combination treatment with 5 μM CBD and zinc ions. The dead cell population (blue rectangles) was increased after combination treatment with CBD and zinc ions (** p ≤ 0.01).

Article Snippet: The expression vectors of MT1G and MT2A were purchased from Origene (RC204741 and RC202748, Rockville, MD, USA).

Techniques: Activity Assay, Concentration Assay, MTT Assay, Flow Cytometry, Staining

Journal: Scientific Reports

Article Title: Metallothionein Genes are Highly Expressed in Malignant Astrocytomas and Associated with Patient Survival

doi: 10.1038/s41598-019-41974-9

Figure Lengend Snippet: MT2 and MT3 gene mRNA expression in different malignancy astrocytomas and Kaplan-Meier survival curves. ( a ) MT2 gene expression level in grade I-III astrocytoma and glioblastoma (GBM) (Mann-Whitney test, *p < 0.050). ( b ) Kaplan-Meier overall survival curves according to low and high MT2 gene expression groups (log-rank test: χ 2 = 8.542, df = 1, p = 0.003). ( c ) MT3 gene expression level in grade I-III astrocytoma and glioblastoma (GBM) (Mann-Whitney test, p > 0.05). ( d ) Kaplan-Meier overall survival curves according to low and high MT3 gene expression groups (log-rank test: χ 2 = 5.694, df = 1, p = 0.017). The middle line on the scatter plot graph represents the median value, whiskers – the lowest and the highest values. Graphs symbols represent different astrocytoma grade: grade I astrocytoma – grey rhombus, grade II astrocytoma – black round, gemistocytic astrocytoma - red dot, grade III astrocytoma – grey triangle, GBM - black rectangular. Gene expression level presented as log 2 (2 −ddCt ).

Article Snippet: Analysis of MT expression in patient age groups have shown that higher MT1A, MT2 and MT3 genes expression level was significantly associated with older patient age (>50 years) (Chi-square test, p < 0.05) (data not shown).

Techniques: Expressing, Gene Expression, MANN-WHITNEY