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目的利用重组质粒pDC316-hIL-24,探讨IL-24对人宫颈癌C33A细胞裸鼠移植瘤的生长及凋亡作用。方法 15只裸鼠腋窝处皮下接种人宫颈癌C33A细胞,建立宫颈癌荷瘤裸鼠模型。随机分为三组,用脂质体包裹pDC316-hIL24重组质粒、pDC316空质粒和磷酸盐缓冲液(PBS),分别于瘤体内注射,比较各组移植瘤生长情况;处死裸鼠剥瘤称重,计算抑瘤率。通过RT-PCR检测IL-24转染至移植瘤内,常规HE染色观察组织形态学变化,免疫组化法检测移植瘤组织中Bax、Bcl-2的表达。结果⑴成功建立移植瘤模型,IL-24基因在C33A细胞中表达,重组质粒组移植瘤生长受到明显抑制,移植瘤体积和瘤重均明显小于空质粒组和PBS组(P<0.05),平均抑瘤率为36%(P<0.05),差异有统计学意义。后两组移植瘤体积和瘤重则无明显差异(P>0.05)。⑵病理学见重组质粒组癌细胞有不同程度的坏死灶,坏死区周围可见凋亡细胞,核碎裂等。⑶免疫组化结果示重组质粒组Bcl-2表达下调,Bax表达上调,于空质粒组和PBS组差异有统计学意义(P<0.05),后两组则无明显差异(P>0.05)。结论重组质粒pDC316-hIL-24对裸鼠人宫颈癌移植瘤的生长具有显著抑制作用,促进肿瘤细胞凋亡,为宫颈癌基因治疗的提供理论依据和实验基础。
Objective To investigate the effect of IL-24 on the growth and apoptosis of transplanted human cervical carcinoma C33A cells in nude mice by using recombinant plasmid pDC316-hIL-24. Methods Fifteen nude mice were subcutaneously injected with C33A cells into the axilla of nude mice to establish a cervical cancer-bearing nude mouse model. The cells were randomly divided into three groups. The pDC316-hIL24 recombinant plasmid, pDC316 empty plasmid and phosphate buffered saline (PBS) were encapsulated by liposomes and injected into the tumor respectively to compare the growth of the transplanted tumor in each group. , Calculate the inhibition rate. The expression of Bax and Bcl-2 in tumor tissue was detected by immunohistochemistry. The expression of IL-24 was detected by RT-PCR. Results (1) The model of xenograft tumor was successfully established. IL-24 gene was expressed in C33A cells. The growth of the transplanted tumor in the recombinant plasmid group was significantly inhibited. The tumor volume and tumor weight were significantly smaller than those in the empty plasmid group and PBS group (P <0.05) The tumor inhibition rate was 36% (P <0.05), the difference was statistically significant. After the two groups of tumor size and tumor weight was no significant difference (P> 0.05). ⑵ Pathology, see the recombinant plasmid group of cancer cells have varying degrees of necrosis, apoptotic cells can be seen around the necrotic area, nuclear fragmentation. (3) The results of immunohistochemistry showed that the expression of Bcl-2 was down-regulated and the expression of Bax was up-regulated in the recombinant plasmid group. There was a significant difference between empty plasmid group and PBS group (P <0.05), but there was no significant difference between the latter two groups (P> 0.05). Conclusion The recombinant plasmid pDC316-hIL-24 can significantly inhibit the growth of human cervical cancer xenografts in nude mice and promote the apoptosis of tumor cells, providing the theoretical basis and experimental basis for the gene therapy of cervical cancer.