Jiang Ninghong, Xie Feng, Chen Limei, Chen Fang, Sui Long
1Medical center for diagnosis and treatment of cervical disease, Obstetrics and Gynecology Hospital, Fudan University, Shanghai, 200011 China.
Shanghai Key Laboratory of Female Reproductive Endocrine Related Diseases, Shanghai, 200011 China.
Infect Agent Cancer. 2020 Feb 17;15:12. doi: 10.1186/s13027-020-0279-9. eCollection 2020.
Cervical cancer is the most common malignancy of the female lower genital tract. In our previous study, we found that TLR4 promotes cervical cancer cell growth in vitro. The aim of this study was to further explore the role of TLR4 in HPV-related cervical cancer in vivo by using a nude mouse xenograft model.
Cervical cancer-derived HeLa and CaSki cells (5 × 10/mL) were either stimulated with an optimal concentration of LPS for the appropriate time (HeLa cells were treated with 1 μg/mL LPS for 1 h, and CaSki cells were treated with 2 μg/mL LPS for 1.5 h) or transfected with TLR4 shRNA and then injected subcutaneously into the dorsal right posterior side of nude mice. The shortest width and longest diameter of the transplanted tumors in the nude mice were measured every 3 days.TLR4, IL-6,iNOS, IL-8,COX-2, MIP-3α, TGF-β1 and VEGF expression levels in the transplanted tumor tissue were detected by immunohistochemistry.
The tumor formation rate was 100% in both HeLa and CaSki nude mouse groups. The tumors grew faster, and the cachexia symptoms were more serious in the LPS groups than in the control group. In contrast, the tumors grew slower, and the cachexia symptoms were milder in the TLR4-silenced groups. TLR4, iNOS, IL-6, MIP-3α and VEGF were highly expressed in the transplanted tumor tissues from the LPS groups, and their expression levels were decreased in the TLR4-silenced groups.
TLR4 expression is closely associated with the tumorigenesis and growth of HPV-positive cervical cancer; TLR4 promotes HPV-positive cervical tumor growth and facilitates the formation of a local immunosuppressive microenvironment. Eventually, these conditions may lead to cervical cancer development.
宫颈癌是女性下生殖道最常见的恶性肿瘤。在我们之前的研究中，我们发现Toll样受体4（TLR4）在体外促进宫颈癌细胞生长。本研究的目的是通过使用裸鼠异种移植模型进一步探讨TLR4在人乳头瘤病毒（HPV）相关宫颈癌体内的作用。
将宫颈癌来源的HeLa和CaSki细胞（5×10⁶/mL）用最佳浓度的脂多糖（LPS）刺激适当时间（HeLa细胞用1μg/mL LPS处理1小时，CaSki细胞用2μg/mL LPS处理1.5小时）或转染TLR4短发夹RNA（shRNA），然后皮下注射到裸鼠的右后背部。每3天测量裸鼠移植瘤的最短宽度和最长直径。通过免疫组织化学检测移植瘤组织中TLR4、白细胞介素-6（IL-6）、诱导型一氧化氮合酶（iNOS）、IL-8、环氧化酶-2（COX-2）、巨噬细胞炎性蛋白-3α（MIP-3α）、转化生长因子-β1（TGF-β1）和血管内皮生长因子（VEGF）的表达水平。
HeLa和CaSki裸鼠组的肿瘤形成率均为100%。LPS组的肿瘤生长更快，恶病质症状比对照组更严重。相比之下，TLR4沉默组的肿瘤生长较慢，恶病质症状较轻。TLR4、iNOS、IL-6、MIP-3α和VEGF在LPS组的移植瘤组织中高表达，而在TLR4沉默组中其表达水平降低。
TLR4表达与HPV阳性宫颈癌的发生和生长密切相关；TLR4促进HPV阳性宫颈肿瘤生长并促进局部免疫抑制微环境的形成。最终，这些情况可能导致宫颈癌的发展。
