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PIK3R1 negatively regulates the epithelial-mesenchymal transition and stem-like phenotype of renal cancer cells through the AKT/GSK3β/CTNNB1 signaling pathway.

Lin Y, Yang Z, Xu A, Dong P, Huang Y, Liu H, Li F, Wang H, Xu Q, Wang Y, Sun D, Zou Y, Zou X, Wang Y, Zhang D, Liu H, Wu X, Zhang M, Fu Y, Cai Z, Liu C, Wu S - Sci Rep (2015)

Bottom Line: The expression of PIK3R1 significantly decreased in pRCC and was further reduced in mRCC compared with normal tissue.Moreover, the PIK3R1 depletion promoted the phosphorylation of AKT in the cells.The knockdown of AKT by shRNA reduced p-GSK3β and CTNNB1 expression in the cells, while the depletion of CTNNB1 impaired stem-like phenotype of the cells.

View Article: PubMed Central - PubMed

Affiliation: Department of Urology, Zhujiang Hospital of Southern Medical University, Guangzhou 510280, China.

ABSTRACT
The phosphoinositide 3-kinase (PI3K)/protein kinase B (AKT) pathway has been identified as an important pathway in renal cell carcinoma (RCC). We have reported a nonsense mutation in PIK3R1, which encodes the regulatory subunit of PI3K, in a metastatic RCC (mRCC), while the mutation was absent in the corresponding primary RCC (pRCC). To identify the function of PIK3R1 in RCC, we examined its expression in normal kidney, pRCC and mRCC by immunohistochemistry and real-time polymerase chain reaction. The expression of PIK3R1 significantly decreased in pRCC and was further reduced in mRCC compared with normal tissue. Besides, its expression levels were negatively correlated with T-category of tumor stage. Additionally, 786-O and A-704 cells with PIK3R1 depletion introduced by CRISPR/Cas9 system displayed enhanced proliferation, migration and epithelial-mesenchymal transition (EMT), and acquired a stem-like phenotype. Moreover, the PIK3R1 depletion promoted the phosphorylation of AKT in the cells. The knockdown of AKT by shRNA reduced p-GSK3β and CTNNB1 expression in the cells, while the depletion of CTNNB1 impaired stem-like phenotype of the cells. Overall, PIK3R1 down-regulation in RCC promotes propagation, migration, EMT and stem-like phenotype in renal cancer cells through the AKT/GSK3β/CTNNB1 pathway, and may contribute to progression and metastasis of RCC.

No MeSH data available.


Related in: MedlinePlus

Downregulation of PIK3R1 correlates with progression and metastasis in RCCs.(a), The representative immunohistochemistry photographs of PIK3R1 expression in normal kidney tissues (n = 13), pRCCs (n = 13) and mRCCs (n = 21). Scale bar = 25 μm. (b), PIK3R1 mRNA levels in the pairs of pRCCs and corresponding normal renal tissues (n = 18). (c), PIK3R1 mRNA expression is negatively correlated with NCAD mRNA expression across a cohort of the pairs of normal renal tissues and pRCCs (n = 18). (d), PIK3R1 mRNA expression in pRCCs according to T category and grade. The TNM cancer staging system was designed to gauge the extent of cancer in a patient's body. T describes the size of the tumor and whether it has invaded nearby tissue, N describes regional lymph nodes that are involved, and M describes distant metastasis (spread of cancer from one body part to another). Grading classification for RCC was based on the Fuhrman grading system. Data are presented as mean ± SD. * P < 0.05, ** P < 0.01.
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f1: Downregulation of PIK3R1 correlates with progression and metastasis in RCCs.(a), The representative immunohistochemistry photographs of PIK3R1 expression in normal kidney tissues (n = 13), pRCCs (n = 13) and mRCCs (n = 21). Scale bar = 25 μm. (b), PIK3R1 mRNA levels in the pairs of pRCCs and corresponding normal renal tissues (n = 18). (c), PIK3R1 mRNA expression is negatively correlated with NCAD mRNA expression across a cohort of the pairs of normal renal tissues and pRCCs (n = 18). (d), PIK3R1 mRNA expression in pRCCs according to T category and grade. The TNM cancer staging system was designed to gauge the extent of cancer in a patient's body. T describes the size of the tumor and whether it has invaded nearby tissue, N describes regional lymph nodes that are involved, and M describes distant metastasis (spread of cancer from one body part to another). Grading classification for RCC was based on the Fuhrman grading system. Data are presented as mean ± SD. * P < 0.05, ** P < 0.01.

Mentions: In order to examine the expression of PIK3R1 in RCC, the protein expression of PIK3R1 in normal kidney (n = 13), pRCC (n = 13) and mRCC (n = 21) was determined by IHC. As shown in Fig. 1a, normal kidney tissues displayed high level of PIK3R1 expression, whereas the expression of PIK3R1 was decreased in pRCC and was further reduced to a lower level in mRCC. The mRNA expression of PIK3R1 was then determined by real-time polymerase chain reaction (RT-PCR). Compared with normal kidney tissue group, the mRNA expression of PIK3R1 was significantly decreased in RCC group (n = 18) (Fig. 1b). The epithelial-mesenchymal transition (EMT) is considered to be crucial to tumor progression and metastasis, in which NCAD is the hallmark of EMT1920. To determine whether the downregulation of PIK3R1 could affect the expression of NCAD, the mRNA expression of NCAD was examined, and data showed that the expression of NCAD had a negative correlation with the mRNA expression of PIK3R1 (Correlation = 0.6929, P = 0.0014) (Fig. 1c). Additionally, the mRNA expression of PIK3R1 negatively correlated with the T category of tumor, although there was no significant difference among different grades (Fig. 1d). These data suggest that the downregulation of PIK3R1 in RCCs correlates with their progression and metastasis.


PIK3R1 negatively regulates the epithelial-mesenchymal transition and stem-like phenotype of renal cancer cells through the AKT/GSK3β/CTNNB1 signaling pathway.

Lin Y, Yang Z, Xu A, Dong P, Huang Y, Liu H, Li F, Wang H, Xu Q, Wang Y, Sun D, Zou Y, Zou X, Wang Y, Zhang D, Liu H, Wu X, Zhang M, Fu Y, Cai Z, Liu C, Wu S - Sci Rep (2015)

Downregulation of PIK3R1 correlates with progression and metastasis in RCCs.(a), The representative immunohistochemistry photographs of PIK3R1 expression in normal kidney tissues (n = 13), pRCCs (n = 13) and mRCCs (n = 21). Scale bar = 25 μm. (b), PIK3R1 mRNA levels in the pairs of pRCCs and corresponding normal renal tissues (n = 18). (c), PIK3R1 mRNA expression is negatively correlated with NCAD mRNA expression across a cohort of the pairs of normal renal tissues and pRCCs (n = 18). (d), PIK3R1 mRNA expression in pRCCs according to T category and grade. The TNM cancer staging system was designed to gauge the extent of cancer in a patient's body. T describes the size of the tumor and whether it has invaded nearby tissue, N describes regional lymph nodes that are involved, and M describes distant metastasis (spread of cancer from one body part to another). Grading classification for RCC was based on the Fuhrman grading system. Data are presented as mean ± SD. * P < 0.05, ** P < 0.01.
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Related In: Results  -  Collection

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getmorefigures.php?uid=PMC4355729&req=5

f1: Downregulation of PIK3R1 correlates with progression and metastasis in RCCs.(a), The representative immunohistochemistry photographs of PIK3R1 expression in normal kidney tissues (n = 13), pRCCs (n = 13) and mRCCs (n = 21). Scale bar = 25 μm. (b), PIK3R1 mRNA levels in the pairs of pRCCs and corresponding normal renal tissues (n = 18). (c), PIK3R1 mRNA expression is negatively correlated with NCAD mRNA expression across a cohort of the pairs of normal renal tissues and pRCCs (n = 18). (d), PIK3R1 mRNA expression in pRCCs according to T category and grade. The TNM cancer staging system was designed to gauge the extent of cancer in a patient's body. T describes the size of the tumor and whether it has invaded nearby tissue, N describes regional lymph nodes that are involved, and M describes distant metastasis (spread of cancer from one body part to another). Grading classification for RCC was based on the Fuhrman grading system. Data are presented as mean ± SD. * P < 0.05, ** P < 0.01.
Mentions: In order to examine the expression of PIK3R1 in RCC, the protein expression of PIK3R1 in normal kidney (n = 13), pRCC (n = 13) and mRCC (n = 21) was determined by IHC. As shown in Fig. 1a, normal kidney tissues displayed high level of PIK3R1 expression, whereas the expression of PIK3R1 was decreased in pRCC and was further reduced to a lower level in mRCC. The mRNA expression of PIK3R1 was then determined by real-time polymerase chain reaction (RT-PCR). Compared with normal kidney tissue group, the mRNA expression of PIK3R1 was significantly decreased in RCC group (n = 18) (Fig. 1b). The epithelial-mesenchymal transition (EMT) is considered to be crucial to tumor progression and metastasis, in which NCAD is the hallmark of EMT1920. To determine whether the downregulation of PIK3R1 could affect the expression of NCAD, the mRNA expression of NCAD was examined, and data showed that the expression of NCAD had a negative correlation with the mRNA expression of PIK3R1 (Correlation = 0.6929, P = 0.0014) (Fig. 1c). Additionally, the mRNA expression of PIK3R1 negatively correlated with the T category of tumor, although there was no significant difference among different grades (Fig. 1d). These data suggest that the downregulation of PIK3R1 in RCCs correlates with their progression and metastasis.

Bottom Line: The expression of PIK3R1 significantly decreased in pRCC and was further reduced in mRCC compared with normal tissue.Moreover, the PIK3R1 depletion promoted the phosphorylation of AKT in the cells.The knockdown of AKT by shRNA reduced p-GSK3β and CTNNB1 expression in the cells, while the depletion of CTNNB1 impaired stem-like phenotype of the cells.

View Article: PubMed Central - PubMed

Affiliation: Department of Urology, Zhujiang Hospital of Southern Medical University, Guangzhou 510280, China.

ABSTRACT
The phosphoinositide 3-kinase (PI3K)/protein kinase B (AKT) pathway has been identified as an important pathway in renal cell carcinoma (RCC). We have reported a nonsense mutation in PIK3R1, which encodes the regulatory subunit of PI3K, in a metastatic RCC (mRCC), while the mutation was absent in the corresponding primary RCC (pRCC). To identify the function of PIK3R1 in RCC, we examined its expression in normal kidney, pRCC and mRCC by immunohistochemistry and real-time polymerase chain reaction. The expression of PIK3R1 significantly decreased in pRCC and was further reduced in mRCC compared with normal tissue. Besides, its expression levels were negatively correlated with T-category of tumor stage. Additionally, 786-O and A-704 cells with PIK3R1 depletion introduced by CRISPR/Cas9 system displayed enhanced proliferation, migration and epithelial-mesenchymal transition (EMT), and acquired a stem-like phenotype. Moreover, the PIK3R1 depletion promoted the phosphorylation of AKT in the cells. The knockdown of AKT by shRNA reduced p-GSK3β and CTNNB1 expression in the cells, while the depletion of CTNNB1 impaired stem-like phenotype of the cells. Overall, PIK3R1 down-regulation in RCC promotes propagation, migration, EMT and stem-like phenotype in renal cancer cells through the AKT/GSK3β/CTNNB1 pathway, and may contribute to progression and metastasis of RCC.

No MeSH data available.


Related in: MedlinePlus