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. 2020 Jan 7;12(1):144.
doi: 10.3390/cancers12010144.

Targeted Nano-Drug Delivery of Colchicine against Colon Cancer Cells by Means of Mesoporous Silica Nanoparticles

Khaled AbouAitah  1   2 , Heba A Hassan  3 , Anna Swiderska-Sroda  1 , Lamiaa Gohar  4 , Olfat G Shaker  5 , Jacek Wojnarowicz  1 , Agnieszka Opalinska  1 , Julita Smalc-Koziorowska  6 , Stanislaw Gierlotka  1 , Witold Lojkowski  1
Affiliations

Targeted Nano-Drug Delivery of Colchicine against Colon Cancer Cells by Means of Mesoporous Silica Nanoparticles

Khaled AbouAitah et al. Cancers (Basel). .

Abstract

Antimitotics are important anticancer agents and include the natural alkaloid prodrug colchicine (COL). However, a major challenge of using COL as an anticancer drug is its cytotoxicity. We developed a novel drug delivery system (DDS) for COL using mesoporous silica nanoparticles (MSNs). The MSNs were functionalized with phosphonate groups, loaded with COL, and coated with folic acid chitosan-glycine complex. The resulting nanoformulation, called MSNsPCOL/CG-FA, was tested for action against cancer and normal cell lines. The anticancer effect was highly enhanced for MSNsPCOL/CG-FA compared to COL. In the case of HCT116 cells, 100% inhibition was achieved. The efficiency of MSNsPCOL/CG-FA ranked in this order: HCT116 (colon cancer) > HepG2 (liver cancer) > PC3 (prostate cancer). MSNsPCOL/CG-FA exhibited low cytotoxicity (4%) compared to COL (~60%) in BJ1 normal cells. The mechanism of action was studied in detail for HCT116 cells and found to be primarily intrinsic apoptosis caused by an enhanced antimitotic effect. Furthermore, a contribution of genetic regulation (metastasis-associated lung adenocarcinoma transcript 1 (MALAT 1), and microRNA (mir-205)) and immunotherapy effects (angiopoietin-2 (Ang-2 protein) and programmed cell death protein 1 (PD-1) was found. Therefore, this study shows enhanced anticancer effects and reduced cytotoxicity of COL with targeted delivery compared to free COL and is a novel method of developing cancer immunotherapy using a low-cost small-molecule natural prodrug.

Keywords: PD-1 immune checkpoint inhibitor and cancer immunotherapy; apoptosis; colchicine alkaloid; colon cancer cells; mesoporous silica nanoparticles; targeted delivery system.

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Conflict of interest statement

The authors declare no conflict of interest.

Figures

Figure 1
Figure 1
Schematic representation of the steps to obtain the proposed drug delivery system with the final product (MSNsPCOL/CG-FA) and various biological evaluations in vitro.
Figure 2
Figure 2
Morphological structures of the materials. (A) Field emission scanning electron microscopy (FE-SEM) of prepared mesoporous silica nanoparticles (MSNs) at different magnifications. (B) High-resolution transmission electron microscopy (HR-TEM) of prepared materials at different stages: Before and after of modification, colchicine (COL) loading, and coating. (C) Scanning transmission electron microscopy (STEM) of prepared materials at different stages: before and after of modification, COL loading, and coating.
Figure 3
Figure 3
Fourier transform infrared spectroscopy (FTIR) spectra of MSNs before and after modification and COL loading, as well as chitosan (C), and glycine (G), folic acid (FA), and COL.
Figure 4
Figure 4
Thermal analysis of materials. (A) Simultaneous thermal analysis (STA) before and after surface modification, COL loading, and coating. (B) DTG analysis before and after surface modification, COL loading, and coating. (C) Differential scanning calorimetry (DSC) analysis before and after surface modification, COL loading, and coating. (D) X-ray diffraction (XRD) analysis before and after surface modification, COL loading, and coating. (E) Zeta potential measurements in aqueous solution before and after surface modification, COL loading, and coating.
Figure 5
Figure 5
In vitro cytotoxicity (as percent inhibition) of MSNs and MSNs functionalized with phosphonate functional groups (MSNsP) for biocompatibility evaluations in cancer and normal cell lines after 24, 48, and 72 h of incubation with cancer cells (liver, HepG2; prostate, PC3; and colon, HCT116) and normal fibroblasts (BJ1). (A) Cytotoxicity of MSNs towards cell lines. (B) Cytotoxicity of MSNsP towards cell lines. Note: A blue asterisk (*) indicates significant (p < 0.05) differences between tested concentrations, whereas an orange asterisk (*) indicates significant differences between cell lines. NS, not significant. All data are expressed as mean ± SD.
Figure 6
Figure 6
In vitro cytotoxicity (as percent inhibition) of the proposed delivery system in cancer and normal cells after 24, 48, and 72 h of incubation with cells. (A) Anticancer effects on HepG2 cancer cells. (B) Anticancer effects on PC3 cancer cells. (C) Anticancer effects on HCT116 cancer cells. (D) Anticancer effects on BJ1 normal cells. Note: A blue asterisk (*) indicates significant (p < 0.05) differences between tested concentrations, whereas an orange asterisk (*) indicates significant differences between tested samples (nanoformulations and COL). NS, not significant. All data are expressed as mean ± SD.
Figure 7
Figure 7
(A) Tubulin inhibition activity of HCT116 cells treated with MSNs and MSNsP as a function of concentration. (B) Tubulin inhibition activity HCT116 cells treated with the proposed delivery system as a function of concentration. (C) Caspase-3 activity of HCT116 cells treated with MSNs and MSNsP as a function of concentration. (D) Caspase-3 activity of HCT116 cells treated with the proposed delivery system and compared to the model anticancer drug (5-FU) as a function of concentration. Note: A blue asterisk (*) indicates significant (p < 0.05) differences between tested concentrations, whereas an orange asterisk (*) indicates significant differences between tested samples. NS, not significant. All data are expressed as mean ± SD.
Figure 8
Figure 8
(A) Bax activity of HCT116 cells treated with MSNs and MSNsP as a function of concentration. (B) Bax activity of HCT116 cells treated with the proposed delivery system and compared to the model anticancer drug (5-FU) as a function of concentration. (C) Bcl-2 inhibition activity of HCT116 cells treated with MSNs and MSNsP as a function of concentration. (D) Bcl-2 inhibition activity of HCT116 cells treated with the proposed delivery system and compared to the model anticancer drug (5-FU) as a function of concentration. Note: A blue asterisk (*) indicates significant (p < 0.05) differences between tested concentrations, whereas an orange asterisk (*) indicates significant differences between tested samples. NS, not significant. All data are expressed as mean ± SD.
Figure 9
Figure 9
(A) BRAF activity of HCT116 cells treated with MSNs and MSNsP as a function of concentration. (B) BRAF activity of HCT116 cells treated with the proposed delivery system and compared to the model anticancer drug (5-FU) as a function of concentration. (C) BRAF inhibition activity of HCT116 cells treated with MSNs and MSNsP as a function of concentration. (D) BRAF inhibition activity of HCT116 cells treated with the proposed delivery system and compared to the model anticancer drug (5-FU) as a function of concentration. Note: A blue asterisk (*) indicates significant (p < 0.05) differences between tested concentrations, whereas an orange asterisk (*) indicates significant differences between tested samples. All data are expressed as mean ± SD.
Figure 10
Figure 10
(A) Cytochrome c triggering of HCT116 cells treated with MSNs and MSNsP at 500 µg/mL. (B) Cytochrome c triggering of HCT116 cells treated at 60 µg/mL with the proposed delivery system for 72 h and compared to the model anticancer drug (5-FU) and control (without any treatment). (C) Mitochondrial membrane potential (Δψm) of HCT116 cells treated with MSNs and MSNsP at 500 µg/mL. (D) Mitochondrial membrane potential of HCT116 cells treated at 60 µg/mL with the proposed delivery system for 72 h and compared to the model anticancer drug (5-FU) and control. Note: An orange asterisk (*) indicates significant (p < 0.05) differences between tested samples. NS, not significant. All data are expressed as mean ± SD.
Figure 11
Figure 11
(A) MALAT-1 protein expression in normal WI-38 cells, HCT116 cells, and HCT116 cells treated with proposed delivery and 5-FU. (B) mir-205 expression in normal WI-38 cells, HCT116 cells, and HCT116 cells treated with proposed delivery and 5-FU. (C) Ang-2 protein expression in normal WI-38 cells, HCT116 cells, and HCT116 cells treated with proposed delivery and 5-FU. (D) CD44 protein expression in normal WI-38 cells, HCT116 cells, and HCT116 cells treated with proposed delivery and 5-FU. (E) PD-1 protein expression in normal WI-38 cells, HCT116 cells, and HCT116 cells treated with proposed delivery and 5-FU. Note: An orange asterisk (*) indicates significant (p < 0.05) differences between tested samples, whereas a blue asterisk (*) indicates significant differences between specific samples. NS, not significant. All data are expressed as mean ± SD.
Figure 12
Figure 12
Schematic representation of cancer cell killing and possible anticancer mechanisms by which the proposed drug delivery system for colchicine (MSNsPCOL/CG-FA nanoformulation) acts in vitro against HCT116 colon cancer cells.
See this image and copyright information in PMC

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