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Targeting sphingosine kinase 2 (SphK2) by ABC294640 inhibits colorectal cancer cell growth in vitro and in vivo.

Xun C, Chen MB, Qi L, Tie-Ning Z, Peng X, Ning L, Zhi-Xiao C, Li-Wei W - J. Exp. Clin. Cancer Res. (2015)

Bottom Line: Moreover, a low concentration of ABC294640 sensitized the activity of 5-FU and cisplatin in vitro.In vivo, ABC294640 oral administration dramatically inhibited HT-29 xenografts growth in nude mice.Targeting of SphK2 by ABC294640 potently inhibits CRC cell growth both in vitro and in vivo, ABC294640 could be developed as a novel therapeutic for the treatment of CRC.

View Article: PubMed Central - PubMed

Affiliation: Department of Oncology, Shanghai General Hospital, Shanghai Jiaotong University, 100 Haining Road, Shanghai, Hongkou District, 200080, China. caixunshanghai2@163.com.

ABSTRACT

Background: Colorectal cancer (CRC) is a major health problem in China and around the world. It is one of the leading causes of cancer-related deaths. Research groups are thus searching for novel and more efficient anti-CRC agents.

Results: Here we demonstrated that ABC294640, a novel SphK2 inhibitor, induced growth inhibition and apoptosis in transformed and primary CRC cells. The SphK activity was remarkably inhibited by ABC294640, accompanied by sphingosine-1-phosphate (S1P) depletion and ceramide incensement in CRC cells. Exogenously-added S1P inhibited ABC294640-induced HT-29 cell lethality. While C6 ceramide and SphK1 inhibitor SKI-II facilitated ABC294640-induced cytotoxicity against HT-29 cells. ABC294640 inhibited AKT-S6K1, but activated JNK signaling in transformed and primary CRC cells. JNK inhibitors (SP600125 and JNKi-II) alleviated ABC294640-induced CRC cell apoptosis. Moreover, a low concentration of ABC294640 sensitized the activity of 5-FU and cisplatin in vitro. In vivo, ABC294640 oral administration dramatically inhibited HT-29 xenografts growth in nude mice.

Conclusions: Targeting of SphK2 by ABC294640 potently inhibits CRC cell growth both in vitro and in vivo, ABC294640 could be developed as a novel therapeutic for the treatment of CRC.

No MeSH data available.


Related in: MedlinePlus

ABC294640 induces growth inhibition and apoptosis in CRC cell lines. The relative growth (vs. Control group) of tested CRC cells with indicated ABC294640 treatment was tested by MTT assay (a and b). HT-29 cells, treated with or without applied ABC294640 for indicated time, were subjected to clonogenicity assay (c), Annexin V FACS assay (d), histone-DNA ELISA assay (e) or LDH release assay (f). Mean values ± SD of three independent experiments were reported. Statistical analysis was performed comparing treatment groups with vehicle control group (“C”). *P < 0.05
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Fig1: ABC294640 induces growth inhibition and apoptosis in CRC cell lines. The relative growth (vs. Control group) of tested CRC cells with indicated ABC294640 treatment was tested by MTT assay (a and b). HT-29 cells, treated with or without applied ABC294640 for indicated time, were subjected to clonogenicity assay (c), Annexin V FACS assay (d), histone-DNA ELISA assay (e) or LDH release assay (f). Mean values ± SD of three independent experiments were reported. Statistical analysis was performed comparing treatment groups with vehicle control group (“C”). *P < 0.05

Mentions: We first examined the potential effect of ABC294640 on CRC cell growth. CRC cell lines, including HT-29, HCT-116 and DLD-1, were treated with applied concentrations of ABC294640, cell growth was tested by MTT assay. Results showed that ABC294640 dose-dependently inhibited CRC cell growth (Fig. 1a). The effect of ABC294640 was also time-dependent, and it took at least 48 hours for ABC294640 to exert the anti-proliferative activity in HT-29 cells (Fig. 1b). Meanwhile, ABC294640 treatment decreased the number of survival HT-29 colonies, further confirming its growth-inhibitory and cytotoxic activities (Fig. 1c). Next, we tested the role of ABC294640 on HT-29 cell apoptosis. Results of Annexin V FACS assay (Fig. 1d) and histone-DNA ELISA assay (Fig. 1e) demonstrated that ABC294640 induced significant apoptosis in HT-29 cells. Meanwhile, LDH content in conditional medium of ABC294640-treated cells was also increased (Fig. 1f). Similar apoptosis and LDH results were also seen in two other CRC cell lines (HCT-116 and DLD-1) (Data not shown). Together, these results show that ABC294640 induces growth inhibition and apoptosis in cultured CRC cells.Fig. 1


Targeting sphingosine kinase 2 (SphK2) by ABC294640 inhibits colorectal cancer cell growth in vitro and in vivo.

Xun C, Chen MB, Qi L, Tie-Ning Z, Peng X, Ning L, Zhi-Xiao C, Li-Wei W - J. Exp. Clin. Cancer Res. (2015)

ABC294640 induces growth inhibition and apoptosis in CRC cell lines. The relative growth (vs. Control group) of tested CRC cells with indicated ABC294640 treatment was tested by MTT assay (a and b). HT-29 cells, treated with or without applied ABC294640 for indicated time, were subjected to clonogenicity assay (c), Annexin V FACS assay (d), histone-DNA ELISA assay (e) or LDH release assay (f). Mean values ± SD of three independent experiments were reported. Statistical analysis was performed comparing treatment groups with vehicle control group (“C”). *P < 0.05
© Copyright Policy - OpenAccess
Related In: Results  -  Collection

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getmorefigures.php?uid=PMC4559903&req=5

Fig1: ABC294640 induces growth inhibition and apoptosis in CRC cell lines. The relative growth (vs. Control group) of tested CRC cells with indicated ABC294640 treatment was tested by MTT assay (a and b). HT-29 cells, treated with or without applied ABC294640 for indicated time, were subjected to clonogenicity assay (c), Annexin V FACS assay (d), histone-DNA ELISA assay (e) or LDH release assay (f). Mean values ± SD of three independent experiments were reported. Statistical analysis was performed comparing treatment groups with vehicle control group (“C”). *P < 0.05
Mentions: We first examined the potential effect of ABC294640 on CRC cell growth. CRC cell lines, including HT-29, HCT-116 and DLD-1, were treated with applied concentrations of ABC294640, cell growth was tested by MTT assay. Results showed that ABC294640 dose-dependently inhibited CRC cell growth (Fig. 1a). The effect of ABC294640 was also time-dependent, and it took at least 48 hours for ABC294640 to exert the anti-proliferative activity in HT-29 cells (Fig. 1b). Meanwhile, ABC294640 treatment decreased the number of survival HT-29 colonies, further confirming its growth-inhibitory and cytotoxic activities (Fig. 1c). Next, we tested the role of ABC294640 on HT-29 cell apoptosis. Results of Annexin V FACS assay (Fig. 1d) and histone-DNA ELISA assay (Fig. 1e) demonstrated that ABC294640 induced significant apoptosis in HT-29 cells. Meanwhile, LDH content in conditional medium of ABC294640-treated cells was also increased (Fig. 1f). Similar apoptosis and LDH results were also seen in two other CRC cell lines (HCT-116 and DLD-1) (Data not shown). Together, these results show that ABC294640 induces growth inhibition and apoptosis in cultured CRC cells.Fig. 1

Bottom Line: Moreover, a low concentration of ABC294640 sensitized the activity of 5-FU and cisplatin in vitro.In vivo, ABC294640 oral administration dramatically inhibited HT-29 xenografts growth in nude mice.Targeting of SphK2 by ABC294640 potently inhibits CRC cell growth both in vitro and in vivo, ABC294640 could be developed as a novel therapeutic for the treatment of CRC.

View Article: PubMed Central - PubMed

Affiliation: Department of Oncology, Shanghai General Hospital, Shanghai Jiaotong University, 100 Haining Road, Shanghai, Hongkou District, 200080, China. caixunshanghai2@163.com.

ABSTRACT

Background: Colorectal cancer (CRC) is a major health problem in China and around the world. It is one of the leading causes of cancer-related deaths. Research groups are thus searching for novel and more efficient anti-CRC agents.

Results: Here we demonstrated that ABC294640, a novel SphK2 inhibitor, induced growth inhibition and apoptosis in transformed and primary CRC cells. The SphK activity was remarkably inhibited by ABC294640, accompanied by sphingosine-1-phosphate (S1P) depletion and ceramide incensement in CRC cells. Exogenously-added S1P inhibited ABC294640-induced HT-29 cell lethality. While C6 ceramide and SphK1 inhibitor SKI-II facilitated ABC294640-induced cytotoxicity against HT-29 cells. ABC294640 inhibited AKT-S6K1, but activated JNK signaling in transformed and primary CRC cells. JNK inhibitors (SP600125 and JNKi-II) alleviated ABC294640-induced CRC cell apoptosis. Moreover, a low concentration of ABC294640 sensitized the activity of 5-FU and cisplatin in vitro. In vivo, ABC294640 oral administration dramatically inhibited HT-29 xenografts growth in nude mice.

Conclusions: Targeting of SphK2 by ABC294640 potently inhibits CRC cell growth both in vitro and in vivo, ABC294640 could be developed as a novel therapeutic for the treatment of CRC.

No MeSH data available.


Related in: MedlinePlus