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. 2020 Jun 17;19(1):52.
doi: 10.1186/s12938-020-00797-w.

An in vitro study on sonodynamic treatment of human colon cancer cells using sinoporphyrin sodium as sonosensitizer

Yuanyuan Shen  1 , Jianquan Ou  1 , Xin Chen  1 , Xiaojun Zeng  2 , Lanhui Huang  2 , Zhaoke Pi  1 , Yaxin Hu  1 , Siping Chen  1 , Tie Chen  3
Affiliations

An in vitro study on sonodynamic treatment of human colon cancer cells using sinoporphyrin sodium as sonosensitizer

Yuanyuan Shen et al. Biomed Eng Online. .

Abstract

Background: Colorectal cancer is the third leading cause of cancer-related deaths worldwide. Sonodynamic therapy (SDT) is an emerging cancer therapy, and in contrast to photodynamic therapy, could non-invasively reach deep-seated tissues and locally activates a sonosensitizer preferentially accumulated in the tumor area to produce cytotoxicity effects. In comparison with traditional treatments, SDT may serve as an alternative strategy for human colon cancer treatment. Here, we investigated the sonodynamic effect using sinoporphyrin sodium (DVDMS) as a novel sonosensitizer on human colon cancer cells in vitro.

Results: The absorption spectra of DVDMS revealed maximum absorption at 363 nm wavelength and emission peak at 635 nm. Confocal microscopy images revealed the DVDMS was primarily localized in the cytoplasm, while no evident signal was detected within the nuclei. Flow cytometry analysis showed rapid intracellular uptake of DVDMS by two types of human colon cancer cells (HCT116 and RKO). Cell viability of HCT116 was tolerant with the concentration of DVDMS up to 20 µg/mL, while the case of RKO was 5 µg/mL. In comparison with the control group, the SDT-treated groups of these two types of human colon cancer cells showed significant increase in cellular apoptosis and necrosis ratio. Increased intracellular reactive oxygen species (ROS) production was detected, indicating the involvement of ROS in mediating SDT effects.

Conclusion: DVDMS results an effective sonosensitizer for the ultrasound-mediated cancer cell killing, and its anticancer effect seems to rely on its ability to produce ROS under ultrasound exposure.

Keywords: Colorectal cancer; Sinoporphyrin sodium; Sonodynamic therapy.

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Conflict of interest statement

The authors declare that they have no competing interests.

Figures

Fig. 1
Fig. 1
Spectral analysis of DVDMS. a Absorption spectra of DVDMS. b Emission spectra of DVDMS
Fig. 2
Fig. 2
Viability of human colon cancer cells and normal colon cells after incubation with DVDMS at different concentrations for 6 h. a HCT116 cells, b RKO cells and c NCM460 cells. *p < 0.05 versus control. **p < 0.01 versus control
Fig. 3
Fig. 3
Intracellular uptake of DVDMS. a Intracellular localization of DVDMS (red) in HCT116 cells and RKO cells. The nuclei were stained with DAPI (blue). Scale bar 50 µm. b Measurement of fluorescence intensity of intracellular DVDMS in HCT116, RKO and NCM460 cells with different incubation durations by a microplate reader and flow cytometry
Fig. 4
Fig. 4
Apoptosis and necrosis analyses of HCT116 cells exposed to different treatment regimens. Representative flow cytometry results of cells at 2 h after exposing to a no treatment, b DVDMS (5 µg/mL) alone, c ultrasound alone, and d SDT using DVDMS (5 µg/mL). e The ratio of apoptotic and necrotic cells in four groups. **p < 0.01 versus control, DVDMS, and US groups
Fig. 5
Fig. 5
Apoptosis and necrosis analyses of RKO cells exposed to different treatment regimens. Representative flow cytometry results of cells at 2 h after exposing to a no treatment, b DVDMS (5 µg/mL) alone, c ultrasound alone, and d SDT using DVDMS (5 µg/mL). e The ratio of apoptotic and necrotic cells in four groups. **p < 0.01 versus control, DVDMS, and US groups
Fig. 6
Fig. 6
Intracellular ROS level of HCT116 and RKO cells in four groups. After SDT treatment, the cells were cultured in the dark for additional 2 h before ROS analyses. a Representative fluorescence images of DCF (green), probing intracellular ROS. b Representative histogram of fluorescence intensity of DCF in HCT116 and RKO cells detected by flow cytometry. Scale bar 100 µm
Fig. 7
Fig. 7
Chemical structure of DVDMS
Fig. 8
Fig. 8
a Graphical illustration of the ultrasonic setup and b acoustic map of peak rarefactional pressure measured at the focal area in the lateral plane
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