Spectral Properties and Antitumor Activity of 10-Hydroxycamptothecin and Its Application in Cell Labeling†

doi:10.7503/cjcu20140575

Abstract

Abstract:

The spectral properties of 10-hydroxycamptothecin(HCPT) were investigated by UV-Vis spectrophotometer, fluorescence spectrophotometer and fourier transform infrared spectroscopy. The antiproliferative activities of HCPT against three tumor cell lines, human cervical can-cer cell line(HeLa), human breast cancer cell line(MCF-7) and human fibrosarcoma cell line(HT1080), were determined using an MTT assay, and the IC₅₀ values of HCPT to the three tumor cell lines were 16.37, 16.73 and 19.24 μg/mL, respectively. The growth inhibitory effect of HCPT on a normal cell line, HEK293T(human embryonic kidney cells 293T), was also investigated, the highest inhibitory rate reached 88.72% at an HCPT concentration of 40 μg/mL, indicating that the normal cell line HEK293T is more sensitive to HCPT in comparison to the three tumor cell lines. Using HeLa cell line as a model, Annexin V-FITC apoptosis detection kit was employed to demonstrate the antitumor mechanism of HCPT. Almost all the cells were simultaneously stained by Annexin V-FITC and PI, the cell membrane was green while the nucleus was red, which indicated that HCPT induced the late apoptosis process of HeLa cells. The distribution of HCPT in HeLa cells was observed by fluorescence microscopy, which provides scientific basis for its application in cell labeling.

Key words: 10-Hydroxycamptothecin, Spectral property, Antitumor activity, Apoptosis, Cell labeling

CLC Number:

O629.3

TrendMD:

DING Guobin, LI Binchun, GUO Yi, XU Li. Spectral Properties and Antitumor Activity of 10-Hydroxycamptothecin and Its Application in Cell Labeling^†[J]. Chem. J. Chinese Universities, 2014, 35(11): 2324.

Figures/Tables 5

Fig.1 UV-Vis spectra of HCPT solutions with different concentrations c/(μg·mL-1): a—g. 1, 5, 10, 20, 30, 40, 50.

Fig.2 Fluorescence spectra of HCPT solutions with different concentrations Inset: the images of HCPT solutions(1, 20 and 50 μg/mL) under an ultraviolet lamp(λex=365 nm).

Fig.3 In vitro antitumor activities of HCPT solutions (A) HCPT solutions with different concentrations against HeLa, MCF-7 and HT1080 cells; (B) growth inhibitory effect of HCPT on HEK293T cells after 48 h incubation at 37 ℃, values are averages of three replicates±SD(standard deviation).

Fig.4 Fluorescence microscopy images of HeLa cells treated with control cell medium(A) and HCPT(40 μg/mL)(B) for 24 h followed by staining with an annexin V-FITC apoptosis detection kit

Fig.5 Fluorescence microscopy images of HeLa cells incubated with control cell medium(A) and HCPT(40 μg/mL)(B) for 12 h

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