Antiviral activity of curcumin-nanoemulsion associated with photodynamic therapy in vulvar cell lines transducing different variants of HPV-16
Antiviral activity of curcumin-nanoemulsion associated with photodynamic therapy in vulvar cell lines transducing different variants of HPV-16
复制标题
DOI：
10.1080/21691401.2020.1725023
复制
发表时间：
2020-01-01
影响因子：
5.8
通讯作者：
Rahal, Paula
中科院分区：
文献类型：
--
作者：
do Bonfim, Caroline Measso;Monteleoni, Leticia Figueiredo;Rahal, Paula
外阴上皮内瘤变（VIN）与人乳头瘤病毒（HPV）感染有关。姜黄素是一种具有抗肿瘤特性的天然生物活性化合物。使用含有姜黄素的纳米颗粒可以使该化合物在治疗中发挥更好的性能。因此，收集了VIN活检并通过PCR进行HPV DNA检测，通过限制性片段长度多态性（RFLP）对阳性样本进行基因分型，并通过测序确定HPV-16变异体。用 E-P 和 E-350G HPV-16 E6 变体转导 HPV-16 阳性外阴癌细胞 (A431)。通过MTT测定测定用纳米乳剂处理的转导细胞的活力。此外，通过Caspase-3/7的酶活性评价细胞凋亡。细胞活力测定表明，与对照细胞相比，空纳米乳液（NE-V）和姜黄素纳米乳液（NE-CUR）对细胞活力的影响很小。此外，我们观察到在 NE-CUR 存在下受辐射的细胞出现 90% 的细胞死亡。细胞凋亡测定进一步显示，用 NE-CUR 处理后，表达 HPV-16 E6 变体的 A431 细胞中 caspase 3 和 7 的活性显着增加。最后，我们将 HPV 转导的 A431 细胞提交到器官型培养物中，并观察到治疗组合会影响组织结构，并产生明显的组织损伤信号。我们得出的结论是，纳米乳液具有良好的生物相容性，因为没有观察到细胞毒性，并且与光活化相关的 NE-CUR 显示出有希望的结果，仅导致受到辐射的细胞死亡。这种与光动力疗法相关的药物输送系统可能在外阴病变的治疗中具有前景。
Vulvar intraepithelial neoplasia (VIN) is associated with human papillomavirus (HPV) infection. Curcumin is a natural bioactive compound with antineoplastic properties. The use of nanoparticles containing curcumin could allow a better performance of this compound in therapies. So, VIN biopsies were collected and HPV DNA detection was performed by PCR, positive samples were genotyped by Restriction Fragment Length Polymorphism (RFLP) and HPV-16 variants were determined by sequencing. HPV-16 positive vulva carcinoma cells (A431) were transduced with E-P and E-350G HPV-16 E6 variants. The viability of the transduced cells treated with nanoemulsions was determined by MTT assay. Besides, apoptosis was evaluated by enzymatic activity of Caspase-3/7. The cell viability assay showed that both the empty nanoemulsion (NE-V) and the nanoemulsion of curcumin (NE-CUR) had little effect on cell viability as compared to control cells. Additionally, we observed that cells irradiated in the presence of NE-CUR presented 90% of cell death. The apoptosis assay further revealed a significant increase in the activity of caspases 3 and 7 in A431 cells expressing both HPV-16 E6 variants after treatment with NE-CUR. Finally, we submitted the HPV transduced A431 cells to organotypic cultures and observed that the combination of treatments affected tissue architecture with evident signals of tissue damage. We concluded that nanoemulsions attain good biocompatibility, since no cytotoxicity was observed and NE-CUR associated with photoactivation showed promising results, leading to death only in cells subjected to irradiation. This drug delivery system associated with photodynamic therapy may become promising in the treatment of vulva lesions.