Korean J Physiol Pharmacol.  2016 May;20(3):253-259. 10.4196/kjpp.2016.20.3.253.

4-(Tert-butyl)-2,6-bis(1-phenylethyl)phenol induces pro-apoptotic activity

• Kim JH
• Lee Y
• Kim MY
• Cho JY

• KMID: 2285597
• DOI: http://doi.org/10.4196/kjpp.2016.20.3.253

Abstract

Previously, we found that KTH-13 isolated from the butanol fraction of Cordyceps bassiana (Cb-BF) displayed anti-cancer activity. To improve its antiproliferative activity and production yield, we employed a total synthetic approach and derivatized KTH-13 to obtain chemical analogs. In this study, one KTH-13 derivative, 4-(tert-butyl)-2,6-bis(1-phenylethyl)phenol (KTH-13-t-Bu), was selected to test its anti-cancer activity. KTH-13-t-Bu diminished the proliferation of C6 glioma, MDA-MB-231, LoVo, and HCT-15 cells. KTH-13-t-Bu induced morphological changes in C6 glioma cells in a dose-dependent manner. KTH-13-t-Bu also increased the level of early apoptotic cells stained with annexin V-FITC. Furthermore, KTH-13-t-Bu increased the levels of cleaved caspase-3 and -9. In contrast, KTH-13-t-Bu upregulated the levels of pro- and cleaved forms of caspase-3, -8, and -9 and Bcl-2. Phospho-STAT3, phospho-Src, and phospho-AKT levels were also diminished by KTH13-t-Bu treatment. Therefore, these results strongly suggest that KTH-13-t-Bu can be considered a novel anti-cancer drug displaying pro-apoptotic activity.

Keyword

Anti-cancer activity; KTH-13-t-Bu; Pro-apoptotic activity; Src; STAT-3

MeSH Terms

Caspase 3
Cordyceps
Glioma
Caspase 3

Figure

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  Fig. 1 Chemical structure of KTH-13-t-Bu.

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  Fig. 2 Effect of KTH-13-t-Bu on proliferation of (A) C6 glioma, (B) MDA-MB-231, (C) LoVo, and (D) HCT-15 cells after treatment for 24 or 6 h.Cell viability was determined by the MTT assay. *p<0.05 and **p<0.01 compared with the normal group.

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  Fig. 3 Effect of KTH-13-t-Bu on inducing apoptosis in C6 glioma cells.(A) C6 glioma cells (5×105 cells/ml) were incubated with KTH-13-t-Bu for 0, 5, and 6 h. Morphological chan ges were detected at each time point by microscopic analysis. (B) Dose-dependent pro-apoptotic effect of KTH-13-t-Bu was measured by FITC annexin V-PI staining assay. Cells were treated with annexin V, PI, and KTH-13-t-Bu (0 to 40 µM) for 6 h. Stained cells were detected by flow cytometry. (C) Changes the nuclear morphology of C6 glioma cells induced by KTH-13-t-Bu were analyzed by confocal microscopy after DAPI staining. (D) Dose-dependent upregulation of apoptosis-related proteins induced by KTH-13-t-Bu. C6 glioma cells (5×105 cells/ml) were incubated with KTH-13-t-Bu for 6 h. The levels of total and cleaved caspase 3, caspase 9, and Bcl-2 were detected by immunoblotting analysis.

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  Fig. 4 Effect of KTH-13-t-Bu on cell survival signaling in C6 glioma cells (5×105 cells/ml).(A and B) STAT3, Src, and AKT phosphorylation were analyzed by immunoblotting analysis.

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  Fig. 5 Putative apoptosis-inducing pathway of KTH-13-t-Bu in cancer cells.

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