S100A9对宫颈鳞癌顺铂化疗敏感性的影响及机制研究

The incidence rates of cervical cancer are highest in gynecological cancers, which threat women's health. Chemo-resistance of cervical cancer cells is an important factor for leading to chemotherapy failure and poor diagnosis. Therefore, to improve the sensitivity of cervical cancer to chemotherapy treatment plays an important role in improving the treatment effects on cervical cancer. In our previous study, we found the expression of S100A9 was associated with carcinogenesis of cervical squamous cancer. In addition, S100A9 protein expression in pre-chemotherapy cancer tissues could predict response to cisplatin-based neoadjuvant chemotherapy in cervical squamous cancer. However, the mechanism of them is still unclear. In recent years, studies reported that chemo-sensitivity of cervical cancer cells to cisplatin was related to many factors including cell apoptosis. In the present study, combined with in vivo and in vitro studies, using construction of recombinant plasmid, RNA interference, flow cytometry, and antibody array, we will explore the effects of S100A9 on malignant characteristic of cervical squamous cancer cells, such as cell apoptosis, cell proliferation, invasion and migration, and investigate the effects of S100A9 on chemo-sensitivity of cervical squamous cancer cells to cisplatin, as well as related role protein and signal transduction pathway. This study will reveal the effects and mechanism of S100A9 on chemo-sensitivity of cervical cancer cells to cisplatin and provide a new thread and experimental basis for improving the chemotherapy effects on cervical cancer.

宫颈癌是最常见的妇科恶性肿瘤之一，严重威胁了女性的健康。宫颈癌化疗耐药是导致化疗失败及影响预后的重要因素，因此提高宫颈癌化疗药物的敏感性是提高宫颈癌治疗效果的关键。在我们的前期研究中，发现S100A9蛋白的表达与宫颈鳞癌的发生相关，同时发现化疗前宫颈鳞癌组织中S100A9蛋白的表达与顺铂为基础的新辅助化疗的敏感性相关，但具体机制尚未明确。近年的研究表明，宫颈癌细胞对顺铂的敏感性与细胞凋亡等多种因素相关。为此，本项目拟结合体内和体外试验，通过质粒转染、RNA干扰、流式细胞仪、抗体芯片等技术，深入研究S100A9对宫颈鳞癌细胞凋亡、增殖、迁移和侵袭等生物学行为的影响；研究S100A9对宫颈鳞癌细胞顺铂化疗药物敏感性的影响及其相关的关键蛋白和信号传导通路，阐明S100A9对宫颈鳞癌顺铂化疗敏感性影响及机制，为提高宫颈癌的化疗效果提供新的思路和实验依据。

宫颈癌是最常见的妇科恶性肿瘤之一，提高宫颈癌化疗药物的敏感性是提高宫颈癌治疗效果的关键。我们的前期研究发现了S100A9蛋白在宫颈上皮组织中的表达随宫颈鳞癌的癌变而增加，同时发现化疗前宫颈鳞癌组织中S100A9蛋白的表达与顺铂为基础的新辅助化疗的敏感性相关，但具体机制和相关信号传导通路尚未明确。本研究结合了体内宫颈癌裸鼠皮下移植瘤和体外细胞试验，通过腺病毒/慢病毒转染、RNA干扰、流式细胞仪等技术，深入研究S100A9及其作用受体AGRE对宫颈鳞癌细胞凋亡、增殖、迁移和侵袭等生物学行为的影响；研究S100A9的表达对宫颈鳞癌细胞顺铂敏感性的影响及其相关的关键蛋白和信号传导通路。.研究结果发现：（1）成功构建S100A9重组腺病毒和慢病毒，成功构建S100A9稳定高表达的人宫颈鳞癌细胞，申报发明专利，并获授权。（2）高表达S100A9的宫颈鳞癌细胞的增殖能力明显增高，S100A9表达下降后，宫颈鳞癌细胞的增殖能力也明显下降。（3）高表达S100A9的宫颈鳞癌细胞的侵袭和迁移能力均明显增高，而S100A9干扰组细胞的穿膜细胞数、穿过小室的细胞数均明显减少，该作用可能与MMPs的表达相关。（4）裸鼠宫颈癌皮下移植瘤实验表明，S100A9高表达组肿瘤明显大，重量也明显增加，而S100A9低表达组肿瘤的大小和重量均明显减小。（5）S100A9在宫颈鳞癌中的表达与高危型HPV16和HPV45相关。（6）S100A9在宫颈鳞癌中的表达与MMP25的表达正相关，而与MMP15和MMP24的表达负相关。（7）S100A9作用受体AGRE的表达会促进宫颈鳞癌细胞的增殖、侵袭和迁移，但抑制细胞凋亡。（8）S100A9通过抑制宫颈鳞癌细胞的凋亡降低对顺铂的敏感性，该作用与Bcl-2，GST-π及LRP相关，可能是通过AKT/ERK/FOXO1信号传导通路起作用的，但具体机制尚待进一步研究。.该研究明确了S100A9及其作用受体AGRE的表达会促进宫颈癌的发生发展，但S100A9是否会促进高危型HPV在宫颈癌变中的作用尚待进一步研究。S100A9的表达降低了宫颈鳞癌细胞对顺铂的敏感性，为开发S100A9抑制剂来提高宫颈鳞癌的顺铂化疗疗效提供思路和实验依据。