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Cystatin C as a p53-inducible apoptotic mediator that regulates cathepsin L activity.

Mori J, Tanikawa C, Funauchi Y, Lo PH, Nakamura Y, Matsuda K - Cancer Sci. (2016)

Bottom Line: We showed that cathepsin L activity was decreased in HCT116 p53(+/+) cells after adriamycin treatment, but not in HCT116 p53(-/-) cells.We also found that knockdown of cystatin C reduced adriamycin-induced caspase-3 activation.Cystatin C expression was significantly downregulated in breast cancer cells with p53 mutations, and decreased cystatin C expression was associated with poor prognosis of breast cancer.

View Article: PubMed Central - PubMed

Affiliation: Laboratory of Molecular Medicine, Human Genome Center, Institute of Medical Science, The University of Tokyo, Minato, Tokyo, Japan.

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Induction of cystatin C by p53. (a, b) Quantitative real‐time PCR (qPCR) analysis of cystatin C in U373MG (p53 mutant) and H1299 (p53 ) cells infected with adenovirus expressing p53 (Ad‐p53) or LacZ (Ad‐LacZ) at an MOI of 10 or 20 (upper panel). β‐actin was used for the normalization of expression levels. Error bars represent SD (n = 3). U373MG and H1299 cells were infected with Ad‐p53 or Ad‐LacZ at MOI of 10 or 20. At 36 h after treatment, whole cell extracts were subjected to Western blotting with anti‐cystatin C, anti‐p21, or anti‐β‐actin antibody (middle). The intensities of protein bands were quantified by densitometry, and the ratio of cystatin C to β‐actin was calculated (lower). (c) Quantitative real‐time PCR analysis of cystatin C expression in the thymus of X‐ray‐irradiated p53−/− or p53+/+ mice (10 Gy) (n = 3 per group). β‐actin was used for the normalization of expression levels. Mice were killed 24 h after irradiation with 10 Gy X‐rays.
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cas12881-fig-0002: Induction of cystatin C by p53. (a, b) Quantitative real‐time PCR (qPCR) analysis of cystatin C in U373MG (p53 mutant) and H1299 (p53 ) cells infected with adenovirus expressing p53 (Ad‐p53) or LacZ (Ad‐LacZ) at an MOI of 10 or 20 (upper panel). β‐actin was used for the normalization of expression levels. Error bars represent SD (n = 3). U373MG and H1299 cells were infected with Ad‐p53 or Ad‐LacZ at MOI of 10 or 20. At 36 h after treatment, whole cell extracts were subjected to Western blotting with anti‐cystatin C, anti‐p21, or anti‐β‐actin antibody (middle). The intensities of protein bands were quantified by densitometry, and the ratio of cystatin C to β‐actin was calculated (lower). (c) Quantitative real‐time PCR analysis of cystatin C expression in the thymus of X‐ray‐irradiated p53−/− or p53+/+ mice (10 Gy) (n = 3 per group). β‐actin was used for the normalization of expression levels. Mice were killed 24 h after irradiation with 10 Gy X‐rays.

Mentions: To further evaluate the induction of cystatin C by p53, U373MG and H1299 cells were infected with Ad‐p53 or Ad‐LacZ. We found that cystatin C mRNA and protein were induced in cells infected with Ad‐p53, indicating the regulation of cystatin C by p53 (Fig. 2a,b).


Cystatin C as a p53-inducible apoptotic mediator that regulates cathepsin L activity.

Mori J, Tanikawa C, Funauchi Y, Lo PH, Nakamura Y, Matsuda K - Cancer Sci. (2016)

Induction of cystatin C by p53. (a, b) Quantitative real‐time PCR (qPCR) analysis of cystatin C in U373MG (p53 mutant) and H1299 (p53 ) cells infected with adenovirus expressing p53 (Ad‐p53) or LacZ (Ad‐LacZ) at an MOI of 10 or 20 (upper panel). β‐actin was used for the normalization of expression levels. Error bars represent SD (n = 3). U373MG and H1299 cells were infected with Ad‐p53 or Ad‐LacZ at MOI of 10 or 20. At 36 h after treatment, whole cell extracts were subjected to Western blotting with anti‐cystatin C, anti‐p21, or anti‐β‐actin antibody (middle). The intensities of protein bands were quantified by densitometry, and the ratio of cystatin C to β‐actin was calculated (lower). (c) Quantitative real‐time PCR analysis of cystatin C expression in the thymus of X‐ray‐irradiated p53−/− or p53+/+ mice (10 Gy) (n = 3 per group). β‐actin was used for the normalization of expression levels. Mice were killed 24 h after irradiation with 10 Gy X‐rays.
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cas12881-fig-0002: Induction of cystatin C by p53. (a, b) Quantitative real‐time PCR (qPCR) analysis of cystatin C in U373MG (p53 mutant) and H1299 (p53 ) cells infected with adenovirus expressing p53 (Ad‐p53) or LacZ (Ad‐LacZ) at an MOI of 10 or 20 (upper panel). β‐actin was used for the normalization of expression levels. Error bars represent SD (n = 3). U373MG and H1299 cells were infected with Ad‐p53 or Ad‐LacZ at MOI of 10 or 20. At 36 h after treatment, whole cell extracts were subjected to Western blotting with anti‐cystatin C, anti‐p21, or anti‐β‐actin antibody (middle). The intensities of protein bands were quantified by densitometry, and the ratio of cystatin C to β‐actin was calculated (lower). (c) Quantitative real‐time PCR analysis of cystatin C expression in the thymus of X‐ray‐irradiated p53−/− or p53+/+ mice (10 Gy) (n = 3 per group). β‐actin was used for the normalization of expression levels. Mice were killed 24 h after irradiation with 10 Gy X‐rays.
Mentions: To further evaluate the induction of cystatin C by p53, U373MG and H1299 cells were infected with Ad‐p53 or Ad‐LacZ. We found that cystatin C mRNA and protein were induced in cells infected with Ad‐p53, indicating the regulation of cystatin C by p53 (Fig. 2a,b).

Bottom Line: We showed that cathepsin L activity was decreased in HCT116 p53(+/+) cells after adriamycin treatment, but not in HCT116 p53(-/-) cells.We also found that knockdown of cystatin C reduced adriamycin-induced caspase-3 activation.Cystatin C expression was significantly downregulated in breast cancer cells with p53 mutations, and decreased cystatin C expression was associated with poor prognosis of breast cancer.

View Article: PubMed Central - PubMed

Affiliation: Laboratory of Molecular Medicine, Human Genome Center, Institute of Medical Science, The University of Tokyo, Minato, Tokyo, Japan.

Show MeSH
Related in: MedlinePlus