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.

中文翻译：

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姜黄素-纳米乳剂的抗病毒活性与光动力疗法在转导HPV-16不同变异的外阴细胞系中的关系。

外阴上皮内瘤变（VIN）与人乳头瘤病毒（HPV）感染有关。姜黄素是具有抗肿瘤作用的天然生物活性化合物。含有姜黄素的纳米颗粒的使用可以使该化合物在治疗中具有更好的性能。因此，收集VIN活检样品并通过PCR进行HPV DNA检测，通过限制性片段长度多态性（RFLP）对阳性样品进行基因分型，并通过测序确定HPV-16变异体。用EP和E-350G HPV-16 E6变体转导HPV-16阳性外阴癌细胞（A431）。通过MTT测定法确定用纳米乳剂处理的转导细胞的活力。此外，通过Caspase-3 / 7的酶促活性评价凋亡。细胞活力测定表明，与对照细胞相比，空的纳米乳液（NE-V）和姜黄素纳米乳液（NE-CUR）对细胞活力的影响均很小。此外，我们观察到在NE-CUR存在下照射的细胞占细胞死亡的90％。凋亡测定进一步揭示了在用NE-CUR处理后表达两种HPV-16 E6变体的A431细胞中胱天蛋白酶3和7的活性显着增加。最后，我们将转导HPV的A431细胞进行器官型培养，并观察到治疗的组合会影响组织结构，并带有明显的组织损伤信号。我们的结论是，由于未观察到细胞毒性，并且与光激活相关的NE-CUR显示出令人鼓舞的结果，因此纳米乳液具有良好的生物相容性，仅在受到辐射的细胞中导致死亡。与光动力疗法相关的这种药物递送系统在外阴病变的治疗中可能变得有希望。