630 nm激光对血卟啉衍生物介导的裸鼠肺腺癌A549移植瘤杀伤效应实验研究

The Experimental Study of Effect and Mechanism of 630-nm Laser on Human Lung Adenocarcinoma Cell Xenograft Model in Nude Mice Mediated by Hematoporphyrin Derivatives

摘要: 目的：探讨630 nm激光对血卟啉衍生物介导的裸鼠肺腺癌移植瘤杀伤效应，为临床肺癌光动力治疗(HPD-PDT)提供理论依据。方法：构建A549人肺腺癌皮下移植瘤裸鼠模型并分组，实验组注射光敏剂后用630 nm激光照射，每2天照射一次，共3次。同时设立空白对照组、单纯光敏剂组、单纯光照组，四组分别于最后一次照射2周后，剥取肿瘤标本，完整取出瘤体，分别计算瘤体体积变化及抑瘤率，对瘤体组织行HE、TUNEL染色，PCR及Western-blot实验。结果：HPD-PDT治疗组肿瘤抑制率明显高于对照组，HE染色显示模型建立成功；TUNEL染色结果显示治疗组凋亡阳性细胞明显增多，凋亡现象明显。PCR检测结果显示与其他三组相比较，光动力治疗组Bcl-2 mRNA的表达下降，而Bax mRNA的表达增高(P < 0.05)，差异均具有统计学意义。Western-blot结果显示光动力治疗组Bcl-2蛋白相对表达下降，而Bax蛋白表达增高(P < 0.05)，差异均具有统计学意义。结论：HPD-PDT治疗能抑制肺腺癌移植瘤细胞的生长增殖，可以诱导肿瘤细胞的凋亡、坏死，进而杀伤肿瘤细胞，对肿瘤组织具有杀伤和抑制作用，表明光动力疗法对于肺癌的治疗有很好的疗效。

Abstract: Objective: To explore the efficacy of photodynamic therapy on the transplantation tumor of lung adeno-carcinoma in mice, and to provide theoretical basis for clinical photodynamic therapy. Methods: The human lung adeno-carcinoma A549 subcutaneous transplantation mouse model was successfully constructed and randomly grouped. The experimental group was injected with photosensitizer and irradiated with a laser of 630 nm. Meanwhile, blank control group, single photosensitizer group and single light group were set up. Photodynamic therapy was given every 2 days and total 3 times. Tumor specimens were removed from the four groups 2 weeks after the last irradiation, and the tumor was completely removed. The tumor volume changes were compared and the tumor inhibition rate was calculated. HE and Tunnel staining, PCR and Western-blot experiments were performed on tumor tissues. Results: The tumor inhibition rate of the HPD-PDT group was significantly higher than that of the other three groups. HE staining pathological sections showed that all tumor cells in the treatment group had different degrees of necrosis. TUNEL staining results showed that most of the cells in the treatment group were expanded, the nuclei were fragmented, and the apoptotic positive cells were significantly increased, with the most obvious apoptosis. PCR results showed that compared with the other three groups, the expression of Bcl-2 mRNA in the photodynamic treatment group decreased, while the expression of Bax mRNA increased (P < 0.05), and the differences were statistically significant. Western-blot results showed that the relative expression of Bcl-2 protein decreased in the photodynamic treatment group, while the expression of Bax protein increased (P < 0.05), and the differences were statistically significant. Conclusion: HPD-PDT can inhibit the growth and proliferation of lung adenocarcinoma cells transplanted tumor, induce the apoptosis and necrosis of tumor cells, and finally kill tumor cells. It has a killing and inhibiting effect on tumor tissues, indicating that photodynamic therapy has a good effect on the treatment of lung cancer.