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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 gene inhibits the proliferation of PC3 cells.(A,B) After transducing the KLK10 gene into the PC3 cell line, the mRNA and protein expression levels of KLK10 were significantly higher in PC3-KLK10 than in PC-3 and the blank Vector group. (C) CCK-8 cell proliferation assay showing the growth deceleration of PC3-KLK10 cell lines 24 h after cell seeding, and this phenomenon lasted for 3 days (P < 0.001). (D) General images of PC3-KLK10- and Vector-transplanted tumours showed that the tumour volume was smaller in the KLK10-over-expressing group than in the Vector group. (E) Tumour volume line graph showed that tumours were successfully generated in nude mice 4 days after cell transplantation; the PC3-KLK10 tumour grew more slowly than the PC3-Vector tumour, and a significant difference in tumour volume was found between these two groups starting from the eighth day. (F) Weight line graph showed that the mouse weight was lower in the PC3-Vector group than in the PC3-KLK10 group from the eighth day, but the difference was not significant (P > 0.05).
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f2: KLK10 gene inhibits the proliferation of PC3 cells.(A,B) After transducing the KLK10 gene into the PC3 cell line, the mRNA and protein expression levels of KLK10 were significantly higher in PC3-KLK10 than in PC-3 and the blank Vector group. (C) CCK-8 cell proliferation assay showing the growth deceleration of PC3-KLK10 cell lines 24 h after cell seeding, and this phenomenon lasted for 3 days (P < 0.001). (D) General images of PC3-KLK10- and Vector-transplanted tumours showed that the tumour volume was smaller in the KLK10-over-expressing group than in the Vector group. (E) Tumour volume line graph showed that tumours were successfully generated in nude mice 4 days after cell transplantation; the PC3-KLK10 tumour grew more slowly than the PC3-Vector tumour, and a significant difference in tumour volume was found between these two groups starting from the eighth day. (F) Weight line graph showed that the mouse weight was lower in the PC3-Vector group than in the PC3-KLK10 group from the eighth day, but the difference was not significant (P > 0.05).

Mentions: Using reconstructed lentiviral vector pLVX-KLK10-IRES-ZsGreen1, viral transfection and cell sorting, a high-purity KLK10-expressed stable cell line PC3-KLK10 was obtained. Its KLK10 expression was much higher than that of the PC3-Vector cell line, as confirmed by quantitative real-time PCR (qRT-PCR) and Western blot (WB) (Fig. 2A,B).


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 proliferation of PC3 cells.(A,B) After transducing the KLK10 gene into the PC3 cell line, the mRNA and protein expression levels of KLK10 were significantly higher in PC3-KLK10 than in PC-3 and the blank Vector group. (C) CCK-8 cell proliferation assay showing the growth deceleration of PC3-KLK10 cell lines 24 h after cell seeding, and this phenomenon lasted for 3 days (P < 0.001). (D) General images of PC3-KLK10- and Vector-transplanted tumours showed that the tumour volume was smaller in the KLK10-over-expressing group than in the Vector group. (E) Tumour volume line graph showed that tumours were successfully generated in nude mice 4 days after cell transplantation; the PC3-KLK10 tumour grew more slowly than the PC3-Vector tumour, and a significant difference in tumour volume was found between these two groups starting from the eighth day. (F) Weight line graph showed that the mouse weight was lower in the PC3-Vector group than in the PC3-KLK10 group from the eighth day, but the difference was not significant (P > 0.05).
© Copyright Policy - open-access
Related In: Results  -  Collection

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

f2: KLK10 gene inhibits the proliferation of PC3 cells.(A,B) After transducing the KLK10 gene into the PC3 cell line, the mRNA and protein expression levels of KLK10 were significantly higher in PC3-KLK10 than in PC-3 and the blank Vector group. (C) CCK-8 cell proliferation assay showing the growth deceleration of PC3-KLK10 cell lines 24 h after cell seeding, and this phenomenon lasted for 3 days (P < 0.001). (D) General images of PC3-KLK10- and Vector-transplanted tumours showed that the tumour volume was smaller in the KLK10-over-expressing group than in the Vector group. (E) Tumour volume line graph showed that tumours were successfully generated in nude mice 4 days after cell transplantation; the PC3-KLK10 tumour grew more slowly than the PC3-Vector tumour, and a significant difference in tumour volume was found between these two groups starting from the eighth day. (F) Weight line graph showed that the mouse weight was lower in the PC3-Vector group than in the PC3-KLK10 group from the eighth day, but the difference was not significant (P > 0.05).
Mentions: Using reconstructed lentiviral vector pLVX-KLK10-IRES-ZsGreen1, viral transfection and cell sorting, a high-purity KLK10-expressed stable cell line PC3-KLK10 was obtained. Its KLK10 expression was much higher than that of the PC3-Vector cell line, as confirmed by quantitative real-time PCR (qRT-PCR) and Western blot (WB) (Fig. 2A,B).

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