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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

18F-FLT Micro PET/CT scan confirmed a low proliferation metabolism in the KLK10-expressing stable PC3 tumour, which is consistent with the low expression of the Ki-67 protein.(A: I, II) 18F-FLT micro PET/CT scan showed that the 18F-FLT uptake of the subcutaneous transplanted tumour was lower in the PC3-KLK10 group (arrow head) than in the Vector group (arrow); in the images, hot spots of 18F-FLT can also be found in the digestive and urinary systems, and the background was clean without the interference of other abnormal hot spots. (B) Bar chart of 18F-FLT uptake semi-analysis showed that the 18F-FLT uptake of the PC3-KLK10 tumour, measured as SUVmean and SUVmax, was significantly lower than that of the PC3-Vector tumour (0.07 ± 0.03 vs. 0.25 ± 0.09; 0.83 ± 0.15 vs. 1.57 ± 0.15)(P < 0.05 and P < 0.01). (C: I-IV) Expression of KLK10 measured by IHC confirmed KLK10 over-expression in the PC3-KLK10-transplanted tumour. (D: I-IV) Expression of the Ki-67 protein measured by IHC showed a significant down-regulation in the PC3-KLK10 group compared with the Vector group.
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f3: 18F-FLT Micro PET/CT scan confirmed a low proliferation metabolism in the KLK10-expressing stable PC3 tumour, which is consistent with the low expression of the Ki-67 protein.(A: I, II) 18F-FLT micro PET/CT scan showed that the 18F-FLT uptake of the subcutaneous transplanted tumour was lower in the PC3-KLK10 group (arrow head) than in the Vector group (arrow); in the images, hot spots of 18F-FLT can also be found in the digestive and urinary systems, and the background was clean without the interference of other abnormal hot spots. (B) Bar chart of 18F-FLT uptake semi-analysis showed that the 18F-FLT uptake of the PC3-KLK10 tumour, measured as SUVmean and SUVmax, was significantly lower than that of the PC3-Vector tumour (0.07 ± 0.03 vs. 0.25 ± 0.09; 0.83 ± 0.15 vs. 1.57 ± 0.15)(P < 0.05 and P < 0.01). (C: I-IV) Expression of KLK10 measured by IHC confirmed KLK10 over-expression in the PC3-KLK10-transplanted tumour. (D: I-IV) Expression of the Ki-67 protein measured by IHC showed a significant down-regulation in the PC3-KLK10 group compared with the Vector group.

Mentions: To non-invasively assess the proliferation of xenograft tumours, 18F-labelled 3′-deoxy-3′-fluorothymidine (18F-FLT) micro positron emission tomography/ computer tomography (PET/CT) scan, using the thymidine analogue 18F-labelled 3′-deoxy-3′-fluorothymidine as imaging agent, was conducted to investigate cellular proliferation in vivo17. 18F-FLT is trapped within the cytosol after being monophosphorylated and enters the exogenous DNA pathway as a specific substrate by the action of thymidine kinase 1 (TK1), a principle enzyme in the salvage pathway of DNA synthesis. Therefore, its accumulation is tightly linked to TK1 enzyme activity and the degree of DNA synthesis18. In our study, hot spots of 18F-FLT were found in subcutaneous tumours and the digestive and urinary systems. The background was clean, without interference from other abnormal hot spots (Fig. 3A). The 18F-FLT uptake of PC3-KLK10 tumour, measured as SUVmean (mean standardised uptake value) and SUVmax (maximal standardised uptake value), was significantly lower than those of PC3-Vector tumour (0.07 ± 0.03 vs. 0.25 ± 0.09; 0.83 ± 0.15 vs. 1.57 ± 0.15) (P < 0.05 and P < 0.01; Fig. 3B), which coincided with the proliferation assays in vitro and in vivo.


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)

18F-FLT Micro PET/CT scan confirmed a low proliferation metabolism in the KLK10-expressing stable PC3 tumour, which is consistent with the low expression of the Ki-67 protein.(A: I, II) 18F-FLT micro PET/CT scan showed that the 18F-FLT uptake of the subcutaneous transplanted tumour was lower in the PC3-KLK10 group (arrow head) than in the Vector group (arrow); in the images, hot spots of 18F-FLT can also be found in the digestive and urinary systems, and the background was clean without the interference of other abnormal hot spots. (B) Bar chart of 18F-FLT uptake semi-analysis showed that the 18F-FLT uptake of the PC3-KLK10 tumour, measured as SUVmean and SUVmax, was significantly lower than that of the PC3-Vector tumour (0.07 ± 0.03 vs. 0.25 ± 0.09; 0.83 ± 0.15 vs. 1.57 ± 0.15)(P < 0.05 and P < 0.01). (C: I-IV) Expression of KLK10 measured by IHC confirmed KLK10 over-expression in the PC3-KLK10-transplanted tumour. (D: I-IV) Expression of the Ki-67 protein measured by IHC showed a significant down-regulation in the PC3-KLK10 group compared with the Vector group.
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f3: 18F-FLT Micro PET/CT scan confirmed a low proliferation metabolism in the KLK10-expressing stable PC3 tumour, which is consistent with the low expression of the Ki-67 protein.(A: I, II) 18F-FLT micro PET/CT scan showed that the 18F-FLT uptake of the subcutaneous transplanted tumour was lower in the PC3-KLK10 group (arrow head) than in the Vector group (arrow); in the images, hot spots of 18F-FLT can also be found in the digestive and urinary systems, and the background was clean without the interference of other abnormal hot spots. (B) Bar chart of 18F-FLT uptake semi-analysis showed that the 18F-FLT uptake of the PC3-KLK10 tumour, measured as SUVmean and SUVmax, was significantly lower than that of the PC3-Vector tumour (0.07 ± 0.03 vs. 0.25 ± 0.09; 0.83 ± 0.15 vs. 1.57 ± 0.15)(P < 0.05 and P < 0.01). (C: I-IV) Expression of KLK10 measured by IHC confirmed KLK10 over-expression in the PC3-KLK10-transplanted tumour. (D: I-IV) Expression of the Ki-67 protein measured by IHC showed a significant down-regulation in the PC3-KLK10 group compared with the Vector group.
Mentions: To non-invasively assess the proliferation of xenograft tumours, 18F-labelled 3′-deoxy-3′-fluorothymidine (18F-FLT) micro positron emission tomography/ computer tomography (PET/CT) scan, using the thymidine analogue 18F-labelled 3′-deoxy-3′-fluorothymidine as imaging agent, was conducted to investigate cellular proliferation in vivo17. 18F-FLT is trapped within the cytosol after being monophosphorylated and enters the exogenous DNA pathway as a specific substrate by the action of thymidine kinase 1 (TK1), a principle enzyme in the salvage pathway of DNA synthesis. Therefore, its accumulation is tightly linked to TK1 enzyme activity and the degree of DNA synthesis18. In our study, hot spots of 18F-FLT were found in subcutaneous tumours and the digestive and urinary systems. The background was clean, without interference from other abnormal hot spots (Fig. 3A). The 18F-FLT uptake of PC3-KLK10 tumour, measured as SUVmean (mean standardised uptake value) and SUVmax (maximal standardised uptake value), was significantly lower than those of PC3-Vector tumour (0.07 ± 0.03 vs. 0.25 ± 0.09; 0.83 ± 0.15 vs. 1.57 ± 0.15) (P < 0.05 and P < 0.01; Fig. 3B), which coincided with the proliferation assays in vitro and in vivo.

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