Journal of Jilin University(Medicine Edition) ›› 2022, Vol. 48 ›› Issue (1): 9-17.doi: 10.13481/j.1671-587X.20220102

• Research in basic medicine • Previous Articles     Next Articles

Establishment of zebrafish xenograft model of nasopharyngeal carcinoma and inhibitory effect of curcumin on CNE-2 cells

Zetai WANG1,Dandan LOU1,Yan PENG1,Daoqi ZHU1,Aiwu LI2,Fengying GONG2,Ying LYU2,Qin FAN1()   

  1. 1.Department of Molecular Biology,School of Traditional Chinese Medicine,Southern Medical University,Guangzhou 510515,China
    2.Traditional Chinese Medicine,NanFang Hospital,Southern Medical University,Guangzhou 510515,China
  • Received:2021-06-04 Online:2022-01-28 Published:2022-01-17
  • Contact: Qin FAN E-mail:fqin@163.com

Abstract: Objective

To establish a zebrafish xenograft model of human nasopharyngeal carcinoma and explore the inhibitory effect of curcumin on nasopharyngeal carcinoma in vivo and in vitro, and to provide the basis for elucidating the molecular mechanism of anti-nasopharyngeal carcinoma of curcumin.

Methods

The nasopharyngeal carcinoma CNE-2 cells were divided into control group and CM-Dil group. The cells were counted for 5 consecutive days, and the fluorescence intensity changes were observed. The zebrafish xenograft model of nasopharyngeal carcinoma was established by microinjection. The zebrafishes were divided into control group, PBS group and model group. The zebrafishes in control group were not treated, the zebrafishes in PBS group were injected with 10 nL PBS buffer, and the zebrafishes in model group were injected with 10 nL cell suspension. The survival number of zebrafishes in each group was counted, and the fluorescence intensity of transplanted tumor of the zebrafishes in model group was observed and quantitatively analyzed by macro stereomicroscope. The morphology of zebrafish transplanted tumors in control group and model group were observed by HE staining. The zebrafishes with the days postfertilization(dpf) was 3 were exposed to feeding water containing different concentrations (0, 0.625, 1.250, 2.500, 5.000,and 7.500, 10.000 μmol·L-1) of curcumin, the death and deformity were observed;the death rates of zebrafishes were calculated,and the concentration of curcumin in vivo was selected. The zebrafish models were treated with differnent concentrations (0,0.625, 1.250, 2.500, and 5.000 μmol·L-1) of curcumin for 48 h, the growth and metastasis of the transplanted tumor were detected. The CNE-2 cells were treated with different concentrations(0, 10, and 20 μmol·L-1) of curcumin for 24 h, the proliferation rates and migration rates were detected by CCK-8 method and wound healing assay.

Results

Compared with control group, there was no significant difference in the number of CNE-2 cells in CM-Dil group (P>0.05). On the 5th day of the experiment, compared with control group, there was no significant difference in the survival number of zebrafishes in PBS group (P>0.05); compared with control group and PBS group, the survival number of zebrafishes in model group was decreased significantly (P<0.01). The red fluorescence intensity in zebrafishes in model group was increased with the prolongation of time. Compared with the first day after injection, the red fluorescence intensity of zebrafishes in model group was increased significantly on the fifth day after injection (P<0.05).The HE staining results showed that the CNE-2 cells were found in the yolk sac of the zebrafishes in model group, indicating that the models were successfully constructed. The zebrafishes in 0.625, 1.250, 2.500, and 5.000 μmol·L-1 concentrations of curcumin groups had no death and obvious toxicity. Compared with control group(0 μmol·L-1 cureumin group),the fluorescence intensities in the zebrafish xenograft tissue in 2.500 and 5.000 μmol·L-1 cureumin groups were signifricantly decreased(P<0.05 or P<0.01), and the number of zebrafishes with head and tail metastasis in 5.00 μmol·L-1 curcumin group was decreased (P<0.05). The results of CCK-8 method and wound healing assay showed that compared with control group (0 μmol·L-1 curcumin group),the proliferation rates and migration rates of CNE-2 cells in 10 and 20 μmol·L-1 curcumin groups were significantly decreased (P<0.05).

Conclusion

The CNE-2 cells can be successfully implanted in the zebrafishes and to estabish the zebrafish models of nasopharyngeal carcinoma. Curcumin can inhibit the proliferation and metastasis of nasopharyngeal carcinoma in vivo and in vitro, and it has a good prospect of cell anti-tumor.

Key words: Curcumin, Nasopharyngeal carcinoma, Xenograft model, Zebrafish, Cell poliferation rate, Cell migration rate

CLC Number: 

  • R285.5