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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

Signaling changes in ABC294640-treated CRC cells. HT-29 cells or primary CRC cells were treated with or without ABC294640 (1/3 μM) for 12 hours, listed proteins were tested by Western blots (a-d). HT-29 cells were pre-treated with SP600125 (5 μM) or JNK inhibitor II (JNKi-II, 5 μM) for 1 hour, followed by ABC294640 (1 μM) stimulation, cell growth (e) and cell apoptosis (f) were analyzed. Mean values ± SD of three independent experiments were reported. Statistical analysis was performed comparing treatment groups with vehicle control group (“C”). *P < 0.05. #P < 0.05 vs. ABC294640 only group (e and f)
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Fig4: Signaling changes in ABC294640-treated CRC cells. HT-29 cells or primary CRC cells were treated with or without ABC294640 (1/3 μM) for 12 hours, listed proteins were tested by Western blots (a-d). HT-29 cells were pre-treated with SP600125 (5 μM) or JNK inhibitor II (JNKi-II, 5 μM) for 1 hour, followed by ABC294640 (1 μM) stimulation, cell growth (e) and cell apoptosis (f) were analyzed. Mean values ± SD of three independent experiments were reported. Statistical analysis was performed comparing treatment groups with vehicle control group (“C”). *P < 0.05. #P < 0.05 vs. ABC294640 only group (e and f)

Mentions: Activation of AKT-mammalian target of rapamycin (mTOR) signaling has been linked to CRC cell survival, proliferation and chemo-resistance [24]. Next, we tested the effect of ABC294640 on AKT-mTOR activation in CRC cells. Activation of AKT was evidence by phosphorylated (p-) AKT at both Ser-473 and Thr-308. Western blot results in Fig. 4a showed that ABC294640 at 1 μM or 3 μM remarkably inhibited AKT phosphorylation at both sites in HT-29 cells. S6K1 phosphorylation, an indicator of mTOR complex C1 (mTORC1) activation, was also dramatically inhibited by ABC294640 (Fig. 4a). Expression of regular AKT and S6K1 was not affected by the same ABC294640 treatment (Fig. 4a). Similar results were also achieved in primary CRC cells (Fig. 4b). Meanwhile, activation of JNK, tested by p-JNK1/2 and p-c-Jun, was induced by same ABC294640 treatment in HT-29 cells (Fig. 4c), and in primary CRC cells (Fig. 4d). Notably, the JNK inhibitors, SP600125 and JNKi-II, suppressed ABC294640-induced HT-29 growth inhibition (Fig. 4e) and apoptosis (Fig. 4f), indicating a pro-apoptotic role of JNK activation by ABC294640 in CRC cells. Similar results were also seen in primary cancer cells (Data not shown).Fig. 4


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)

Signaling changes in ABC294640-treated CRC cells. HT-29 cells or primary CRC cells were treated with or without ABC294640 (1/3 μM) for 12 hours, listed proteins were tested by Western blots (a-d). HT-29 cells were pre-treated with SP600125 (5 μM) or JNK inhibitor II (JNKi-II, 5 μM) for 1 hour, followed by ABC294640 (1 μM) stimulation, cell growth (e) and cell apoptosis (f) were analyzed. Mean values ± SD of three independent experiments were reported. Statistical analysis was performed comparing treatment groups with vehicle control group (“C”). *P < 0.05. #P < 0.05 vs. ABC294640 only group (e and f)
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Fig4: Signaling changes in ABC294640-treated CRC cells. HT-29 cells or primary CRC cells were treated with or without ABC294640 (1/3 μM) for 12 hours, listed proteins were tested by Western blots (a-d). HT-29 cells were pre-treated with SP600125 (5 μM) or JNK inhibitor II (JNKi-II, 5 μM) for 1 hour, followed by ABC294640 (1 μM) stimulation, cell growth (e) and cell apoptosis (f) were analyzed. Mean values ± SD of three independent experiments were reported. Statistical analysis was performed comparing treatment groups with vehicle control group (“C”). *P < 0.05. #P < 0.05 vs. ABC294640 only group (e and f)
Mentions: Activation of AKT-mammalian target of rapamycin (mTOR) signaling has been linked to CRC cell survival, proliferation and chemo-resistance [24]. Next, we tested the effect of ABC294640 on AKT-mTOR activation in CRC cells. Activation of AKT was evidence by phosphorylated (p-) AKT at both Ser-473 and Thr-308. Western blot results in Fig. 4a showed that ABC294640 at 1 μM or 3 μM remarkably inhibited AKT phosphorylation at both sites in HT-29 cells. S6K1 phosphorylation, an indicator of mTOR complex C1 (mTORC1) activation, was also dramatically inhibited by ABC294640 (Fig. 4a). Expression of regular AKT and S6K1 was not affected by the same ABC294640 treatment (Fig. 4a). Similar results were also achieved in primary CRC cells (Fig. 4b). Meanwhile, activation of JNK, tested by p-JNK1/2 and p-c-Jun, was induced by same ABC294640 treatment in HT-29 cells (Fig. 4c), and in primary CRC cells (Fig. 4d). Notably, the JNK inhibitors, SP600125 and JNKi-II, suppressed ABC294640-induced HT-29 growth inhibition (Fig. 4e) and apoptosis (Fig. 4f), indicating a pro-apoptotic role of JNK activation by ABC294640 in CRC cells. Similar results were also seen in primary cancer cells (Data not shown).Fig. 4

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