Caspase-3对5-ALA诱导鳞状细胞癌Colo-16细胞凋亡效应的影响

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Abstract Objective To investigate the role of caspase-3 in the apoptosis of squamous-cell carcinoma Colo-16 cells induced by 5-ALA-PDT.Methods Colo-16 cells in the logarithmic growth phase were divided into three groups: control group, 5-ALA photodynamic therapy (PDT) group, and caspase-3 inhibitor group (Z-DEVD-FMK). Control group was not given photosensitizer or light treatment. 5-ALA PDT group was exposed to 3-minute laser irradiation at a power density of 10 mW/cm2 and energy density of 2.5 J/cm2 after 4 hours of dark incubation with 0.4 mg/ml 5-ALA before being cultured for 12 hours. Caspase3 inhibitor group was treated in the same way as 5-ALA PDT group except that Z-DEVD-FMK at a final concentration of 40 μmol was added during incubation. Cells in each group were collected after experiment, the changes of intracellular caspase-3 fluorescence intensity were observed by immunofluorescence, and the percentage of positive cells and apoptosis rate of active caspase-3 were detected by flow cytometry.Results Immunofluorescence Results showed that there was weak green fluorescence in the cytoplasm of Colo-16 cells in control group and Caspase-3 inhibitor group, indicating a low expression of caspase-3, while green fluorescence was significantly enhanced in 5-ALA-PDT group, showing a significant increase of caspase-3 content in cytoplasm of Colo-16 cells. Flow cytometry Results showed that the respective percentage of active caspase-3 positive cells was (2.26±0.16)％,(32.16±1.31)％ and (3.36±0.31)％ in control group, 5-ALA PDT group and caspase-3 inhibitor group respectively. The difference was statistically significant (P<0.05). The apoptosis rate of control group, 5-ALA PDT group and caspase-3 inhibitor group was (2.35±0.22)％,(31.55±3.68)％ and (11.46±2.25)％, respectively. Paired comparison was made between the three groups, and the differences were statistically significant (P<0.01).Conclusions The apoptosis of Colo-16 cells induced by 5-ALA-PDT is closely related to the activation of caspase-3.

Key words： caspase-3 human cutaneous carcinoma cell line Colo-16 5-ALA-PDT apoptosis

Received: 20 October 2016

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	Articles by authors
	XU Jin
	QIAO Li
	YANG Zhiyong
	and LIU Wei

Cite this article:

XU Jin,QIAO Li,YANG Zhiyong等. Effects of caspase 3 on apoptosis of squamous-cell carcinoma Colo-16 cells induced by 5-ALA-PDT[J]. Med. J. Chin. Peop. Armed Poli. Forc., 2017, 28(3): 232-234.

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http://journal08.magtechjournal.com/Jwk_wjyx/EN/ OR http://journal08.magtechjournal.com/Jwk_wjyx/EN/Y2017/V28/I3/232

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