Limits...
NES1/KLK10 gene represses proliferation, enhances apoptosis and down-regulates glucose metabolism of PC3 prostate cancer cells.

Hu J, Lei H, Fei X, Liang S, Xu H, Qin D, Wang Y, Wu Y, Li B - Sci Rep (2015)

Bottom Line: Furthermore, by up-regulating Bcl-2 or HK-2 respectively in the PC3-KLK10 cell line, we observed a subsequent increase of cell proliferation and a synchronous up-regulation of HK-2 and Bcl-2.Besides, KLK10 expression was also increased by Bcl-2 and HK-2, which suggests that there is a negative feedback loop between KLK10 and Bcl-2/HK-2.Thus, our results demonstrated that KLK10 may function as a tumour suppressor by repressing proliferation, enhancing apoptosis and decreasing glucose metabolism in PC3 cells.

View Article: PubMed Central - PubMed

Affiliation: Department of Nuclear Medicine, Rui jin Hospital, School of Medicine, Shanghai JiaoTong University; 197 Ruijin Second Road, Shanghai 200025, China.

ABSTRACT
The normal epithelial cell-specific-1 (NES1) gene, also named as KLK10, is recognised as a novel putative tumour suppressor in breast cancer, but few studies have focused on the function of KLK10 in human prostate cancer. Our study confirms that the expression of KLK10 in prostate cancer tissue and cell lines (PC3, DU145, and LNCaP clone FGC) is low. Given that the androgen-independent growth characteristic of the PC3 cell line is more similar to clinical castration-resistant prostate cancer, we studied the role of KLK10 in PC3. In vitro and in vivo assays showed that over-expressing KLK10 in PC3 could decelerate tumour proliferation, which was accompanied with an increase in apoptosis and suppression of glucose metabolism. The related proteins, such as Bcl-2 and HK-2, were down-regulated subsequently. Furthermore, by up-regulating Bcl-2 or HK-2 respectively in the PC3-KLK10 cell line, we observed a subsequent increase of cell proliferation and a synchronous up-regulation of HK-2 and Bcl-2. Besides, KLK10 expression was also increased by Bcl-2 and HK-2, which suggests that there is a negative feedback loop between KLK10 and Bcl-2/HK-2. Thus, our results demonstrated that KLK10 may function as a tumour suppressor by repressing proliferation, enhancing apoptosis and decreasing glucose metabolism in PC3 cells.

No MeSH data available.


Related in: MedlinePlus

KLK10 gene inhibits the glucose metabolism of PC3 cells with the lower 18F-FDG uptake via the down-regulation of HK-2.(A: I, II) 18F-FDG micro PET/CT scan showed that the18F-FDG uptake of the transplanted tumour was lower in the PC3-KLK10 group (arrow head) than in the Vector group (arrow); in the images, hot spots of 18F-FDG could also be found in several organs such as the heart, liver, spleen, digestive and urinary system, but these hot spots did not disturb the imaging of the subcutaneous transplantation tumour. (B) Bar chart of 18F-FDG uptake semi-analysis showed that the uptake of 18F-FDG in KLK10-over-expressing PC3 transplantation tumour was low, with low SUVmean and SUVmax (0.39 ± 0.05 vs. 0.74 ± 0.16, 0.58 ± 0.07 vs. 1.09 ± 0.18; P < 0.05, P < 0.01). (C) The mRNA of HK-2 was much lower in the PC3-KLK10 cell line (P < 0.001) than in the control group. (D) WB showed that the HK-2 protein was significantly down-regulated in the PC3-KLK10 cell line. (E: I-IV) Low expression of the HK-2 protein in the PC3-KLK10 xenograft tumour tissue measured by IHC confirmed the result of WB.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4663538&req=5

f5: KLK10 gene inhibits the glucose metabolism of PC3 cells with the lower 18F-FDG uptake via the down-regulation of HK-2.(A: I, II) 18F-FDG micro PET/CT scan showed that the18F-FDG uptake of the transplanted tumour was lower in the PC3-KLK10 group (arrow head) than in the Vector group (arrow); in the images, hot spots of 18F-FDG could also be found in several organs such as the heart, liver, spleen, digestive and urinary system, but these hot spots did not disturb the imaging of the subcutaneous transplantation tumour. (B) Bar chart of 18F-FDG uptake semi-analysis showed that the uptake of 18F-FDG in KLK10-over-expressing PC3 transplantation tumour was low, with low SUVmean and SUVmax (0.39 ± 0.05 vs. 0.74 ± 0.16, 0.58 ± 0.07 vs. 1.09 ± 0.18; P < 0.05, P < 0.01). (C) The mRNA of HK-2 was much lower in the PC3-KLK10 cell line (P < 0.001) than in the control group. (D) WB showed that the HK-2 protein was significantly down-regulated in the PC3-KLK10 cell line. (E: I-IV) Low expression of the HK-2 protein in the PC3-KLK10 xenograft tumour tissue measured by IHC confirmed the result of WB.

Mentions: The low apoptotic index of PC3-KLK10 might not be the only reason to explain the deceleration of cell proliferation. To satisfy the demand of tumour proliferation and metastasis, cancer cells need an increase in glucose consumption because of the high rates of glycolysis, termed as ‘the Warburg effect’19. Enhanced glucose uptake is sufficiently prevalent such that it is taken advantage of to image cancer cells in clinical application using the glucose analogue 18F labelled 2-fluoro-2-deoxy-D-glucose (18F-FDG) by PET or PET/CT, which is available for diagnosing and staging cancer, evaluating treatment and monitoring cancer relapse for patients20. In our study, we utilised 18F-FDG micro PET/CT scan to mimic the clinical situation for monitoring the effect of KLK10 gene therapy on prostate cancer in vivo. As expected, the glucose metabolism of PC3-KLK10 xenograft tumour was lower than that of the PC3-Vector group. In nude mice, several organs, such as the heart, liver and spleen, could uptake high levels of 18F-FDG, without disturbing the imaging of the subcutaneous transplantation tumour (Fig. 5A). The uptake of 18F-FDG in PC3-KLK10 xenograft tumour was much lower than that in the Vector group, with lower SUVmean and SUVmax (0.39 ± 0.05 vs. 0.74 ± 0.16 and 0.58 ± 0.07 vs. 1.09 ± 0.18, respectively; P < 0.05 and P < 0.01, respectively; Fig. 5B). As a result, the effect of proliferative inhibition of KLK10 in the PC3 cell line might be the combined result of apoptotic induction and glucose metabolism reduction.


NES1/KLK10 gene represses proliferation, enhances apoptosis and down-regulates glucose metabolism of PC3 prostate cancer cells.

Hu J, Lei H, Fei X, Liang S, Xu H, Qin D, Wang Y, Wu Y, Li B - Sci Rep (2015)

KLK10 gene inhibits the glucose metabolism of PC3 cells with the lower 18F-FDG uptake via the down-regulation of HK-2.(A: I, II) 18F-FDG micro PET/CT scan showed that the18F-FDG uptake of the transplanted tumour was lower in the PC3-KLK10 group (arrow head) than in the Vector group (arrow); in the images, hot spots of 18F-FDG could also be found in several organs such as the heart, liver, spleen, digestive and urinary system, but these hot spots did not disturb the imaging of the subcutaneous transplantation tumour. (B) Bar chart of 18F-FDG uptake semi-analysis showed that the uptake of 18F-FDG in KLK10-over-expressing PC3 transplantation tumour was low, with low SUVmean and SUVmax (0.39 ± 0.05 vs. 0.74 ± 0.16, 0.58 ± 0.07 vs. 1.09 ± 0.18; P < 0.05, P < 0.01). (C) The mRNA of HK-2 was much lower in the PC3-KLK10 cell line (P < 0.001) than in the control group. (D) WB showed that the HK-2 protein was significantly down-regulated in the PC3-KLK10 cell line. (E: I-IV) Low expression of the HK-2 protein in the PC3-KLK10 xenograft tumour tissue measured by IHC confirmed the result of WB.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4663538&req=5

f5: KLK10 gene inhibits the glucose metabolism of PC3 cells with the lower 18F-FDG uptake via the down-regulation of HK-2.(A: I, II) 18F-FDG micro PET/CT scan showed that the18F-FDG uptake of the transplanted tumour was lower in the PC3-KLK10 group (arrow head) than in the Vector group (arrow); in the images, hot spots of 18F-FDG could also be found in several organs such as the heart, liver, spleen, digestive and urinary system, but these hot spots did not disturb the imaging of the subcutaneous transplantation tumour. (B) Bar chart of 18F-FDG uptake semi-analysis showed that the uptake of 18F-FDG in KLK10-over-expressing PC3 transplantation tumour was low, with low SUVmean and SUVmax (0.39 ± 0.05 vs. 0.74 ± 0.16, 0.58 ± 0.07 vs. 1.09 ± 0.18; P < 0.05, P < 0.01). (C) The mRNA of HK-2 was much lower in the PC3-KLK10 cell line (P < 0.001) than in the control group. (D) WB showed that the HK-2 protein was significantly down-regulated in the PC3-KLK10 cell line. (E: I-IV) Low expression of the HK-2 protein in the PC3-KLK10 xenograft tumour tissue measured by IHC confirmed the result of WB.
Mentions: The low apoptotic index of PC3-KLK10 might not be the only reason to explain the deceleration of cell proliferation. To satisfy the demand of tumour proliferation and metastasis, cancer cells need an increase in glucose consumption because of the high rates of glycolysis, termed as ‘the Warburg effect’19. Enhanced glucose uptake is sufficiently prevalent such that it is taken advantage of to image cancer cells in clinical application using the glucose analogue 18F labelled 2-fluoro-2-deoxy-D-glucose (18F-FDG) by PET or PET/CT, which is available for diagnosing and staging cancer, evaluating treatment and monitoring cancer relapse for patients20. In our study, we utilised 18F-FDG micro PET/CT scan to mimic the clinical situation for monitoring the effect of KLK10 gene therapy on prostate cancer in vivo. As expected, the glucose metabolism of PC3-KLK10 xenograft tumour was lower than that of the PC3-Vector group. In nude mice, several organs, such as the heart, liver and spleen, could uptake high levels of 18F-FDG, without disturbing the imaging of the subcutaneous transplantation tumour (Fig. 5A). The uptake of 18F-FDG in PC3-KLK10 xenograft tumour was much lower than that in the Vector group, with lower SUVmean and SUVmax (0.39 ± 0.05 vs. 0.74 ± 0.16 and 0.58 ± 0.07 vs. 1.09 ± 0.18, respectively; P < 0.05 and P < 0.01, respectively; Fig. 5B). As a result, the effect of proliferative inhibition of KLK10 in the PC3 cell line might be the combined result of apoptotic induction and glucose metabolism reduction.

Bottom Line: Furthermore, by up-regulating Bcl-2 or HK-2 respectively in the PC3-KLK10 cell line, we observed a subsequent increase of cell proliferation and a synchronous up-regulation of HK-2 and Bcl-2.Besides, KLK10 expression was also increased by Bcl-2 and HK-2, which suggests that there is a negative feedback loop between KLK10 and Bcl-2/HK-2.Thus, our results demonstrated that KLK10 may function as a tumour suppressor by repressing proliferation, enhancing apoptosis and decreasing glucose metabolism in PC3 cells.

View Article: PubMed Central - PubMed

Affiliation: Department of Nuclear Medicine, Rui jin Hospital, School of Medicine, Shanghai JiaoTong University; 197 Ruijin Second Road, Shanghai 200025, China.

ABSTRACT
The normal epithelial cell-specific-1 (NES1) gene, also named as KLK10, is recognised as a novel putative tumour suppressor in breast cancer, but few studies have focused on the function of KLK10 in human prostate cancer. Our study confirms that the expression of KLK10 in prostate cancer tissue and cell lines (PC3, DU145, and LNCaP clone FGC) is low. Given that the androgen-independent growth characteristic of the PC3 cell line is more similar to clinical castration-resistant prostate cancer, we studied the role of KLK10 in PC3. In vitro and in vivo assays showed that over-expressing KLK10 in PC3 could decelerate tumour proliferation, which was accompanied with an increase in apoptosis and suppression of glucose metabolism. The related proteins, such as Bcl-2 and HK-2, were down-regulated subsequently. Furthermore, by up-regulating Bcl-2 or HK-2 respectively in the PC3-KLK10 cell line, we observed a subsequent increase of cell proliferation and a synchronous up-regulation of HK-2 and Bcl-2. Besides, KLK10 expression was also increased by Bcl-2 and HK-2, which suggests that there is a negative feedback loop between KLK10 and Bcl-2/HK-2. Thus, our results demonstrated that KLK10 may function as a tumour suppressor by repressing proliferation, enhancing apoptosis and decreasing glucose metabolism in PC3 cells.

No MeSH data available.


Related in: MedlinePlus