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目的:研究串珠素(perlecan)反义cDNA质粒(pAP)对喉癌Hep-2细胞裸鼠移植瘤的影响。方法:应用pAP转染喉癌Hep-2细胞,建立人喉癌裸鼠移植瘤模型,观察裸鼠移植瘤的生长速度。分为3组:未转染的Hep-2细胞组(WT组)、空载体phβApr-neol转染组(neo组)及pAP转染组(pAP组)。用RT-PCR和免疫组织化学方法检测各组裸鼠移植瘤的perlecan mRNA和蛋白质的表达情况。结果:肿瘤生长4周后,WT组和neo组裸鼠移植瘤的平均体积较大,pAP组裸鼠移植瘤的平均体积较小,差异有统计学意义(P<0.01)。perlecan mRNA在WT组和neo组高表达,在pAP组低表达,均差异有统计学意义(均P<0.05)。perlecan蛋白在WT组和neo组高表达,定位于癌细胞的细胞核和细胞质,在pAP组低表达,均差异有统计学意义(均P<0.01)。结论:pAP对喉癌Hep-2细胞裸鼠移植瘤的发生、发展有重要影响。
Objective: To study the effect of perlecan antisense cDNA plasmid (pAP) on the transplanted tumor of Hep-2 cells in nude mice. Methods: The laryngeal carcinoma Hep-2 cells were transfected with pAP to establish a human laryngeal carcinoma xenograft model in nude mice. The growth rate of nude mice xenografts was observed. The cells were divided into 3 groups: untransfected Hep-2 cells (WT group), empty vector phβApr-neol transfected group (neo group) and pAP transfected group (pAP group). The expression of perlecan mRNA and protein in transplanted xenografts in nude mice was detected by RT-PCR and immunohistochemistry. Results: After 4 weeks of tumor growth, the average volume of xenografts in nude mice in WT group and neo group was larger than that in nude mice in pAP group. The difference was statistically significant (P <0.01). Perlecan mRNA was highly expressed in WT group and neo group, but low in pAP group (all P <0.05). Perlecan protein was highly expressed in WT and neo groups and localized in the nucleus and cytoplasm of cancer cells. The expression of perlecan protein was low in pAP group (all P <0.01). Conclusion: pAP plays an important role in the occurrence and development of transplanted tumor of Hep-2 cells in nude mice.