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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

KLK10-induced apoptosis of PC3 cells results in the accumulation of PC3 cells in sub-G1 phase with the down-regulation of Bcl-2.(A: I-II) Cell cycle showed a significant accumulation of PC3-KLK10 cells in the sub-G1 phase, indicating apoptosis, compared with PC3-Vector cells (9.01% ± 3.165% vs. 0.82% ± 0.372%, P < 0.05). The percentage of PC3-KLK10 cells in the G1 phase was significantly reduced (50.74% ± 3.446% vs. 67.41 ± 4.69%, P < 0.01). (B: I-III) TUNEL assay staining of apoptotic cells showed obvious positive cells with brown colouring in PC3-KLK10 cell nuclei (arrow head). The apoptotic index of PC3-KLK10 was 14.76% ± 1.950%, which was higher than that of PC3-Vector cells (4.36% ± 1.031%)(P < 0.0001). (C) qRT-PCR showed the significant down-regulation of Bcl-2 and the slight down-regulation of Bax in the PC3-KLK10 cell line (P < 0.05 and P > 0.05). (D) WB showed the significant down-regulation of the Bcl-2 and Bax proteins in KLK10-over-expressing PC3 cells. (E: I-IV) Low expression of the Bcl-2 protein in PC3-KLK10 xenograft tumour tissue measured by IHC confirmed the result of WB.
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f4: KLK10-induced apoptosis of PC3 cells results in the accumulation of PC3 cells in sub-G1 phase with the down-regulation of Bcl-2.(A: I-II) Cell cycle showed a significant accumulation of PC3-KLK10 cells in the sub-G1 phase, indicating apoptosis, compared with PC3-Vector cells (9.01% ± 3.165% vs. 0.82% ± 0.372%, P < 0.05). The percentage of PC3-KLK10 cells in the G1 phase was significantly reduced (50.74% ± 3.446% vs. 67.41 ± 4.69%, P < 0.01). (B: I-III) TUNEL assay staining of apoptotic cells showed obvious positive cells with brown colouring in PC3-KLK10 cell nuclei (arrow head). The apoptotic index of PC3-KLK10 was 14.76% ± 1.950%, which was higher than that of PC3-Vector cells (4.36% ± 1.031%)(P < 0.0001). (C) qRT-PCR showed the significant down-regulation of Bcl-2 and the slight down-regulation of Bax in the PC3-KLK10 cell line (P < 0.05 and P > 0.05). (D) WB showed the significant down-regulation of the Bcl-2 and Bax proteins in KLK10-over-expressing PC3 cells. (E: I-IV) Low expression of the Bcl-2 protein in PC3-KLK10 xenograft tumour tissue measured by IHC confirmed the result of WB.

Mentions: To test the underlying mechanism that leads to the KLK10-induced deceleration of cell proliferation, we observed the effects of KLK10 on PC3 cells by detecting cell cycle progression and apoptosis. A significant accumulation of PC3-KLK10 cells in the sub-G1 phase, indicative of apoptosis, was observed compared with PC3-Vector cells (9.01% ± 3.165% vs. 0.82% ± 0.372%; P < 0.05; Fig. 4A). The percentage of PC3-KLK10 cells was significantly reduced in the G1 phase (50.74% ± 3.446% vs. 67.41% ± 4.69%, P < 0.01) but slightly increased in the S phase and G2/M phase without a significant difference (P > 0.05; Fig. 4A). Terminal deoxynucleotidyl transferase-mediated dUTP nick end labelling staining of apoptotic cells showed obvious positive brown cells in PC3-KLK10 cell nuclei (Fig. 4B-II). The apoptotic index of PC3-KLK10 was 14.76% ± 1.950%, which was more than that of PC3-Vector cells (4.36% ± 1.031%; P < 0.001; Fig. 4B-III). The results showed that KLK10 could promote apoptosis of PC3 cells. Although the percentage of apoptosis in PC3-KLK10 cells was not very high, this phenomenon may explain the deceleration of cell proliferation.


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-induced apoptosis of PC3 cells results in the accumulation of PC3 cells in sub-G1 phase with the down-regulation of Bcl-2.(A: I-II) Cell cycle showed a significant accumulation of PC3-KLK10 cells in the sub-G1 phase, indicating apoptosis, compared with PC3-Vector cells (9.01% ± 3.165% vs. 0.82% ± 0.372%, P < 0.05). The percentage of PC3-KLK10 cells in the G1 phase was significantly reduced (50.74% ± 3.446% vs. 67.41 ± 4.69%, P < 0.01). (B: I-III) TUNEL assay staining of apoptotic cells showed obvious positive cells with brown colouring in PC3-KLK10 cell nuclei (arrow head). The apoptotic index of PC3-KLK10 was 14.76% ± 1.950%, which was higher than that of PC3-Vector cells (4.36% ± 1.031%)(P < 0.0001). (C) qRT-PCR showed the significant down-regulation of Bcl-2 and the slight down-regulation of Bax in the PC3-KLK10 cell line (P < 0.05 and P > 0.05). (D) WB showed the significant down-regulation of the Bcl-2 and Bax proteins in KLK10-over-expressing PC3 cells. (E: I-IV) Low expression of the Bcl-2 protein in PC3-KLK10 xenograft tumour tissue measured by IHC confirmed the result of WB.
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f4: KLK10-induced apoptosis of PC3 cells results in the accumulation of PC3 cells in sub-G1 phase with the down-regulation of Bcl-2.(A: I-II) Cell cycle showed a significant accumulation of PC3-KLK10 cells in the sub-G1 phase, indicating apoptosis, compared with PC3-Vector cells (9.01% ± 3.165% vs. 0.82% ± 0.372%, P < 0.05). The percentage of PC3-KLK10 cells in the G1 phase was significantly reduced (50.74% ± 3.446% vs. 67.41 ± 4.69%, P < 0.01). (B: I-III) TUNEL assay staining of apoptotic cells showed obvious positive cells with brown colouring in PC3-KLK10 cell nuclei (arrow head). The apoptotic index of PC3-KLK10 was 14.76% ± 1.950%, which was higher than that of PC3-Vector cells (4.36% ± 1.031%)(P < 0.0001). (C) qRT-PCR showed the significant down-regulation of Bcl-2 and the slight down-regulation of Bax in the PC3-KLK10 cell line (P < 0.05 and P > 0.05). (D) WB showed the significant down-regulation of the Bcl-2 and Bax proteins in KLK10-over-expressing PC3 cells. (E: I-IV) Low expression of the Bcl-2 protein in PC3-KLK10 xenograft tumour tissue measured by IHC confirmed the result of WB.
Mentions: To test the underlying mechanism that leads to the KLK10-induced deceleration of cell proliferation, we observed the effects of KLK10 on PC3 cells by detecting cell cycle progression and apoptosis. A significant accumulation of PC3-KLK10 cells in the sub-G1 phase, indicative of apoptosis, was observed compared with PC3-Vector cells (9.01% ± 3.165% vs. 0.82% ± 0.372%; P < 0.05; Fig. 4A). The percentage of PC3-KLK10 cells was significantly reduced in the G1 phase (50.74% ± 3.446% vs. 67.41% ± 4.69%, P < 0.01) but slightly increased in the S phase and G2/M phase without a significant difference (P > 0.05; Fig. 4A). Terminal deoxynucleotidyl transferase-mediated dUTP nick end labelling staining of apoptotic cells showed obvious positive brown cells in PC3-KLK10 cell nuclei (Fig. 4B-II). The apoptotic index of PC3-KLK10 was 14.76% ± 1.950%, which was more than that of PC3-Vector cells (4.36% ± 1.031%; P < 0.001; Fig. 4B-III). The results showed that KLK10 could promote apoptosis of PC3 cells. Although the percentage of apoptosis in PC3-KLK10 cells was not very high, this phenomenon may explain the deceleration of cell proliferation.

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