Raw data for graphs.

Objective
To investigate the anti-cancer efficacy of ENB101-LNP, an ionizable lipid nanoparticles (LNPs) encapsulating siRNA against E6/E7 of HPV 16, in combination therapy with cisplatin in cervical cancer in vitro and in vivo.
Methods
CaSki cells were treated with ENB101-LNP, cisplatin, or combination. Cell viability assessed the cytotoxicity of the treatment. HPV16 E6/E7 gene knockdown was verified with RT-PCR both in vitro and in vivo. HLA class I and PD-L1 were checked by flow cytometry. A xenograft model was made using CaSki cells in BALB/c nude mice. To evaluate anticancer efficacy, mice were grouped. ENB101-LNP was given three times weekly for 3 weeks intravenously, and cisplatin was given once weekly intraperitoneally. Tumor growth was monitored. On day 25, mice were euthanized; tumors were collected, weighed, and imaged. Tumor samples were analyzed through histopathology, immunostaining, and western blot.
Results
ENB101-LNP and cisplatin synergistically inhibit CaSki cell growth. The combination reduces HPV 16 E6/E7 mRNA and boosts p21 mRNA, p53, p21, and HLA class I proteins. In mice, the treatment significantly blocked tumor growth and promoted apoptosis. Tumor inhibition rates were 29.7% (1 mpk ENB101-LNP), 29.6% (3 mpk), 34.0% (cisplatin), 47.0% (1 mpk ENB101-LNP-cisplatin), and 68.8% (3 mpk ENB101-LNP-cisplatin). RT-PCR confirmed up to 80% knockdown of HPV16 E6/E7 in the ENB101-LNP groups. Immunohistochemistry revealed increased p53, p21, and HLA-A expression with ENB101-LNP treatments, alone or combined.
Conclusion
The combination of ENB101-LNP, which inhibits E6/E7 of HPV 16, with cisplatin, demonstrated significant anticancer activity in the xenograft mouse model of cervical cancer.

研究目标：探讨可电离脂质纳米颗粒（ionizable lipid nanoparticles, LNPs）ENB101-LNP（包裹针对人乳头瘤病毒16型（Human Papillomavirus 16, HPV 16）E6/E7靶点的小干扰RNA（small interfering RNA, siRNA））与顺铂联合疗法在体内外对宫颈癌的抗癌活性。
实验方法：将CaSki细胞分别用ENB101-LNP、顺铂或二者联合处理，通过细胞活力实验评估该处理的细胞毒性；采用实时荧光定量聚合酶链式反应（reverse transcription-polymerase chain reaction, RT-PCR）在体内外验证HPV 16 E6/E7基因的敲低效果；通过流式细胞术检测人类白细胞抗原I类（Human Leukocyte Antigen class I, HLA class I）分子与程序性死亡受体配体1（programmed death-ligand 1, PD-L1）的表达水平。使用CaSki细胞构建BALB/c裸鼠异种移植瘤模型，将荷瘤小鼠随机分组以评估抗癌活性：尾静脉注射给予ENB101-LNP，每周3次，连续给药3周；腹腔注射给予顺铂，每周1次。监测肿瘤生长情况，于第25天时处死小鼠，收集肿瘤组织并称重、成像；通过组织病理学、免疫染色及蛋白质印迹（western blot）分析肿瘤样本。
实验结果：ENB101-LNP与顺铂可协同抑制CaSki细胞增殖。联合给药可降低HPV 16 E6/E7的mRNA水平，并上调p21 mRNA及p53、p21、HLA class I蛋白的表达。在荷瘤小鼠模型中，该治疗方案可显著阻滞肿瘤生长并诱导细胞凋亡。各组肿瘤抑制率分别为：1 mg/kg（mpk）ENB101-LNP组29.7%、3 mg/kg（mpk）ENB101-LNP组29.6%、顺铂单药组34.0%、1 mg/kg（mpk）ENB101-LNP联合顺铂组47.0%、3 mg/kg（mpk）ENB101-LNP联合顺铂组68.8%。RT-PCR检测证实，ENB101-LNP单药组的HPV 16 E6/E7基因敲低效率可达80%。免疫组化结果显示，单用或联合使用ENB101-LNP均可上调p53、p21及人类白细胞抗原A（Human Leukocyte Antigen A, HLA-A）的表达水平。
研究结论：靶向抑制HPV 16 E6/E7的ENB101-LNP与顺铂联合疗法，在宫颈癌异种移植瘤小鼠模型中展现出显著的抗癌活性。