Journal: Journal of Cellular and Molecular Medicine

Article Title: Molecular analysis of sunitinib resistant renal cell carcinoma cells after sequential treatment with RAD001 (everolimus) or sorafenib

doi: 10.1111/jcmm.12471

Figure Lengend Snippet: (A) Cell growth analysis of Caki-1, KTC-26 and A498 cells treated short-term (1 day) or long-term (8 weeks) with 1 μM sunitinib. Controls remained untreated. The figure shows one representative from six separate experiments. *indicates significant difference to controls. (B) Cell cycle analysis of Caki-1, KTC-26 and A498 cells treated short-term (1 day) or long-term (8 weeks) with 1 μM sunitinib. The cell population at each specific checkpoint is expressed as percentage of the total cells analysed. – indicates controls + indicates sunitinib treatment. One representative experiment of three is shown.

Article Snippet: A498 cells were derived from Cell Lines Service (Heidelberg, Germany).

Techniques: Cell Cycle Assay

Journal: Journal of Cellular and Molecular Medicine

Article Title: Molecular analysis of sunitinib resistant renal cell carcinoma cells after sequential treatment with RAD001 (everolimus) or sorafenib

doi: 10.1111/jcmm.12471

Figure Lengend Snippet: (Upper) Evaluation of tumour cell growth of sunitinib-resistant tumour cells exposed to sorafenib. Sunitinib resistant Caki-1, KTC-26 or A498 cells were treated short-term (1 day) or long-term (8 weeks) with 5 μM sorafenib (sunitinib-sorafenib). Controls were treated with culture medium alone (medium-control). To define the maximum effect of sorafenib, tumour cells not pre-treated with sunitinib were exposed to sorafenib (sorafenib-control). The figure shows one representative from six separate experiments. *indicates significant difference to controls. (Lower) Modulation of cell cycle progression in second line setting. Sunitinib resistant Caki-1, KTC-26 or A498 cells were treated short-term (1 day) or long-term (8 weeks) with 5 μM sorafenib (suni-sora). Controls were treated with culture medium alone (medium). To define the maximum effect of sorafenib, tumour cells not pre-treated with sunitinib were exposed to sorafenib (sorafenib). The cell population at each specific checkpoint is expressed as percentage of the total cells analysed. One representative experiment of three is shown.

Article Snippet: A498 cells were derived from Cell Lines Service (Heidelberg, Germany).

Techniques: Control

Journal: Journal of Cellular and Molecular Medicine

Article Title: Molecular analysis of sunitinib resistant renal cell carcinoma cells after sequential treatment with RAD001 (everolimus) or sorafenib

doi: 10.1111/jcmm.12471

Figure Lengend Snippet: (Upper) Evaluation of tumour cell growth of sunitinib-resistant tumour cells exposed to RAD001. Sunitinib resistant Caki-1, KTC-26 or A498 cells were treated short-term (1 day) or long-term (8 weeks) with 5 nM RAD001 (sunitinib-RAD001). Controls were treated with culture medium alone (medium-control). To define the maximum effect of RAD001, tumour cells not pre-treated with sunitinib were exposed to RAD001 (RAD001-control). The figure shows one representative from six separate experiments. *indicates significant difference to controls. (Lower) Modulation of cell cycle progression in second line setting. Sunitinib resistant Caki-1, KTC-26 or A498 cells were treated short-term (1 day) or long-term (8 weeks) with 5 nM RAD001 (suni-RAD). Controls were treated with culture medium alone (medium). To define the maximum effect of RAD001, tumour cells not pre-treated with sunitinib were exposed to RAD001 (RAD001). The cell population at each specific checkpoint is expressed as percentage of the total cells analysed. One representative experiment of three is shown.

Article Snippet: A498 cells were derived from Cell Lines Service (Heidelberg, Germany).

Techniques: Control

Journal: Journal of Cellular and Molecular Medicine

Article Title: Molecular analysis of sunitinib resistant renal cell carcinoma cells after sequential treatment with RAD001 (everolimus) or sorafenib

doi: 10.1111/jcmm.12471

Figure Lengend Snippet: Analysis of cell cycle regulating proteins in Caki-1, KTC-26 and A498 cells after switching to sorafenib versus RAD001 treatment. Sunitinib resistant RCC cells were treated short-term (1 day) or long-term (8 weeks) with 5 μM sorafenib (suni+sora) or 5 nM RAD001 (suni+RAD001). Controls were treated with culture medium alone (control). To define the maximum effect of sorafenib and RAD001, tumour cells not pre-treated with sunitinib were exposed to sorafenib (5 μM; sorafenib) or RAD001 (5 nM; RAD001). Sunitinib resistant cells, which were further treated with sunitinib for 1 day or 8 weeks were also subjected to the assay (sunitinib). β-actin served as the internal control. The figure shows one representative from three separate experiments.

Article Snippet: A498 cells were derived from Cell Lines Service (Heidelberg, Germany).

Techniques: Control

Journal: Cell Death & Disease

Article Title: REGγ deficiency suppresses tumor progression via stabilizing CK1ε in renal cell carcinoma

doi: 10.1038/s41419-018-0646-2

Figure Lengend Snippet: a , b Expression of REGγ in RCC tissues (T) and normal kidney tissues (N) as detected by IHC ( a ) and WB ( b ). c Expression of REGγ in four RCC cell lines (A498, 786-O, ACHN, and caki-1) and the normal renal tubular epithelial cell line (HK-2) as detected by western blot. d Comparison of REGγ expression in different Fuhrman grades I–IV of RCC tissue samples via IHC staining (left panel: magnification ×200, scale bar = 50 μm; right panel: magnification ×400, scale bar = 20 μm). e REGγ (PSME3) expression (median of expression intensity) in different pathological types and grades of RCC derived from Oncomine database ( https://www.oncomine.org/ ). f Kaplan–Meier analysis of the correlation between REGγ expression and the survival time in RCC patients. Cases were classified into lower expression group and higher expression group as described in methods. g Prognosis of RCC (KIRC) patients with high or low expression of REGγ (PSME3) derived from OncoLnc database ( https://www.oncolnc.org/ )

Article Snippet: The stable REGγ knockdown RCC cell lines ACHN and A498 were generated by integration of retroviral shREGγ vectors specific for REGγ or a control gene (GFP) from OriGene (Rockville, MD), which was performed as previously described .

Techniques: Expressing, Western Blot, Immunohistochemistry, Derivative Assay

Journal: Cell Death & Disease

Article Title: REGγ deficiency suppresses tumor progression via stabilizing CK1ε in renal cell carcinoma

doi: 10.1038/s41419-018-0646-2

Figure Lengend Snippet: a PSME3 upregulation is closely correlated with renal cell carcinoma revealed by KEGG pathway analysis. b GSEA indicated that cell proliferation and anti-apoptosis were positively correlated with elevated PSME3 expression in RCC from database GSE89563 (GO_0008284 and GO_0006915). NES, normalized enrichment score. c Stable knockdown of REGγ (shREGγ) in RCC cell lines (ACHN and A498) confirmed by WB. d , e Effect of shREGγ on RCC cells growth as determined by MTT assay. f Representative images of colony formation of RCC cells after transfection of shREGγ versus shNC. g , h Representative images of EdU incorporation assay after transfection of shREGγ versus shNC. i , j Representative flow cytometry plots of cell cycle distribution from ACHN and A498 cells transfected with shREGγ and shNC. k , l Apoptosis rate from ACHN and A498 cells after transfected with shREGγ and shNC as detected by flow cytometry. Data are shown as mean ± SD. * P < 0.05

Article Snippet: The stable REGγ knockdown RCC cell lines ACHN and A498 were generated by integration of retroviral shREGγ vectors specific for REGγ or a control gene (GFP) from OriGene (Rockville, MD), which was performed as previously described .

Techniques: Expressing, MTT Assay, Transfection, Flow Cytometry

Journal: Cell Death & Disease

Article Title: REGγ deficiency suppresses tumor progression via stabilizing CK1ε in renal cell carcinoma

doi: 10.1038/s41419-018-0646-2

Figure Lengend Snippet: a , b Representative images and the relative quantification of wound-healing assay in RCC cells transfected with shREGγ and shNC. c , d Representative images and the relative quantification of transwell invasion assay in ACHN and A498 cells transfected with shREGγ and shNC. Data are shown as mean ± SD. * P < 0.05

Article Snippet: The stable REGγ knockdown RCC cell lines ACHN and A498 were generated by integration of retroviral shREGγ vectors specific for REGγ or a control gene (GFP) from OriGene (Rockville, MD), which was performed as previously described .

Techniques: Wound Healing Assay, Transfection, Transwell Invasion Assay

Journal: Scientific Reports

Article Title: ORM1 promotes tumor progression of kidney renal clear cell carcinoma (KIRC) through CALR-mediated apoptosis

doi: 10.1038/s41598-023-42962-w

Figure Lengend Snippet: ORM1 was essential to cell proliferation. ( a ) The expression of ORM1 protein in A498, 786-O, and Caki-2 cells was much higher than the 293 T cells. ( b ) The gray value analysis of protein in ( a ). ( c ) ORM1 was knocked down in 786-O and Caki-2 cells. ( d ) The gray value analysis of protein in ( c ). ( e ) Cell proliferation of 786-O cells was inhibited in ORM1-KD group compared to NC group. ( f ) Cell proliferation of Caki-2 cells was inhibited in ORM1-KD group compared to NC group. # p < 0.05 showed statistically difference.

Article Snippet: KIRC cell lines including 786-O, A498, and Caki-2 cells were obtained from ICell Bioscience Inc, Shanghai (Shanghai, China).

Techniques: Expressing

Journal: Scientific Reports

Article Title: ORM1 promotes tumor progression of kidney renal clear cell carcinoma (KIRC) through CALR-mediated apoptosis

doi: 10.1038/s41598-023-42962-w

Figure Lengend Snippet: ORM1 was essential to cell migration and invasion. ( a ) Cell migration and invasion was suppressed in ORM1-KD group compared to NC group in 786-O and Caki-2 cells in transwell assay with/without Matrigel. ( b ) The statistical analysis of cells in cell migration in ( a ). ( c ) The statistical analysis of cells in cell invasion in a. # p < 0.05 showed statistically difference.

Article Snippet: KIRC cell lines including 786-O, A498, and Caki-2 cells were obtained from ICell Bioscience Inc, Shanghai (Shanghai, China).

Techniques: Migration, Transwell Assay

Journal: Scientific Reports

Article Title: ORM1 promotes tumor progression of kidney renal clear cell carcinoma (KIRC) through CALR-mediated apoptosis

doi: 10.1038/s41598-023-42962-w

Figure Lengend Snippet: ORM1 enhanced the efficiency of sorafenib in KIRC. ( a ) Sorafenib inhibited cell proliferation in concentration-dependent manner. ( b ) The efficiency of sorafenib was enhanced in ORM1-KD group but relieved in ORM1-OE group in 786-O cells. ( c ) The efficiency of sorafenib was enhanced in ORM1-KD group but relieved in ORM1-OE group in Caki-2 cells. # p < 0.05 showed statistically difference.

Article Snippet: KIRC cell lines including 786-O, A498, and Caki-2 cells were obtained from ICell Bioscience Inc, Shanghai (Shanghai, China).

Techniques: Concentration Assay