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. 2015 Dec;64(5):473-479.
doi: 10.7727/wimj.2016.053. Epub 2016 Apr 29.

The Effect and Mechanisms of Proliferative Inhibition of Crocin on Human Leukaemia Jurkat Cells

Y Sun  1 , Z Wang  2 , L Wang  3 , L-Z Wang  1 , C Zang  4 , L-R Sun  5
Affiliations

The Effect and Mechanisms of Proliferative Inhibition of Crocin on Human Leukaemia Jurkat Cells

Y Sun et al. West Indian Med J. 2015 Dec.

Abstract
in English, Spanish

Targeted therapy is a potentially useful approach for the treatment of T-lineage acute lymphoblastic leukaemia. This study aimed to find a highly effective, low toxic anti-tumour drug and further investigate its mechanisms. Jurkat cells were used as the object and were stimulated by different concentrations of crocin. By cell count, growth curve, MTT method for the detection of cell proliferation, annexin V/propidium iodide (PI) method for the apoptosis rates, and reverse transcription-polymerase chain reaction (RT-PCR) for bcl-2 and bax gene expression, the effect and mechanisms of proliferative inhibition of crocin on Jurkat cells were further explored. Crocin promoted Jurkat cell apoptosis and inhibited cell growth, in a dose-time-dependent manner. The mechanism might be related to the inhibition of bcl-2 gene expression and the promotion of bax gene expression. These results suggest that crocin can be used as a suitable clinical agent for the treatment of T-lineage acute lymphoblastic leukaemia.

La terapia dirigida es un método potencialmente útil para el tratamiento de la leucemia linfoblástica aguda de linaje T. Este estudio tuvo por objetivo encontrar un medicamento antitumoral de baja toxicidad y altamente eficaz, contra esta enfermedad, y seguir investigando sus mecanismos. Las células Jurkat fueron utilizadas como objeto y se les estimuló con diferentes concentraciones de crocina. El efecto y los mecanismos de inhibición proliferativa de crocina en células Jurkat fueron además explorados mediante recuento de células, curva de crecimiento, método metil tiazolil tetrazolio (MTT) para la detección de la proliferación celular, método anexina V/PI para las tasas de apoptosis, y la reacción en cadena reversa de la transcriptasa-polimerasa (RT-PCR) para la expresión del gen Bcl-2 y el gen Bax. La crocina promueve la apoptosis de las células Jurkat e inhibe el crecimiento celular, de manera dosistiempo dependiente. El mecanismo podría estar relacionado con la inhibición de la expresión del gen Bcl-2 y la promoción de la expresión del gen Bax. Estos resultados sugieren que la crocina puede utilizarse como agente clínico adecuado para el tratamiento de la leucemia linfoblástica aguda de linaje T.

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Figures

Fig. 1
Fig. 1. The morphology of Jurkat cells cultured for 48 hours. The cells of the control group showed round contours, transparent and uniform cytoplasm, good refraction and appearance of a bunch of grapes in suspension. With the crocin concentration of 5 mg/mL and 10 mg/mL, cells showed shrunken body, weakened refraction, increasing intracellular particles and a single suspended growth.
Fig. 2
Fig. 2. Cell count at different times and different concentrations of crocin.
Fig. 3
Fig. 3. The apoptotic rates of cells at different times and different concentrations of crocin.
Fig. 4
Fig. 4. Expression of Bcl-2 gene at different concentrations of crocin. M: Marker; Lanes 7–12: GAPDH; Lane 1: 0 mg/mL; Lane 2: 0.625 mg/mL; Lane 3: 1.25 mg/mL; Lane 4: 2.5mg/mL; Lane 5: 5 mg/mL; Lane 6: 10 mg/mL
Fig. 5
Fig. 5. Expression of Bax gene at different concentrations of crocin. M: Marker; Lanes 7–12: GAPDH; Lane 1: 0 mg/mL; Lane 2: 0.625 mg/mL; Lane 3: 1.25 mg/mL; Lane 4: 2.5 mg/mL; Lane 5: 5 mg/mL; Lane 6:10 mg/mL.
Fig. 6
Fig. 6. Apoptosis and gene expression. (1) bcl-2 and apoptosis; (2) bax and apoptosis; (3) bcl-2 and bax.
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