Department of Hepatobiliary and Echinococcosis Surgery, Digestive and Vascular Surgery Center, First Affiliated Hospital of Xinjiang Medical University, 830054, Urumqi, Xinjiang, China; State Key Laboratory of Pathogenesis, Prevention and Treatment of High Incidence Diseases in Central Asia, Xinjiang Medical University, 830054, Urumqi, Xinjiang, China.
Department of Hepatobiliary and Echinococcosis Surgery, Digestive and Vascular Surgery Center, First Affiliated Hospital of Xinjiang Medical University, 830054, Urumqi, Xinjiang, China.
Exp Parasitol. 2022 Sep;240:108321. doi: 10.1016/j.exppara.2022.108321. Epub 2022 Jul 2.
Immune cells are pivotal players in the immune responses against both parasitic infection and malignancies. Substantial evidence demonstrated that there may exist possible relationship between echinococcus granulus sensu lato (E. granulosus s.l.) infection and hepatocellular carcinoma (HCC) development. Thus, this study aimed to observe crucial roles of immune cells in the formation of subcutaneous lesions after transplanting HepG2 cell lines with or without E. granulosus s.l. protoscoleces (PSCs). HepG2 cell lines were subcutaneously injected into nude mice in the control group. In the co-transplantation group, HepG2 cells were subcutaneously co-injected with high dosage of E. granulosus s.l. PSCs. From the 25th day of transplantation, volume of subcutaneous lesions was measured every four days, which were removed at the 37th day for further studies. Basic pathological and functional changes were observed. Moreover, expression of Ki67, Bcl-2, Caspase3, α-smooth muscle actin (α-SMA), T cell markers (CD3, CD4, CD8), PD1/PD-L1, nature killer (NK) cell markers (CD16, CD56) were further detected by immunohistochemical staining and quantitative real-time polymerase chain reaction (qRT-PCR) analysis. Subcutaneous lesions were gradually increased in volume and there occurred pathologically heterogeneous tumor cells, which were more significant in the co-transplantation group. Compared to the control group, expression of proliferation markers Ki67 and Bcl-2 was at higher levels in the co-transplantation group. Reversely, apoptotic marker Caspase3 was highly detected in the control group, suggesting promoting effects of E. granulosus s.l. PSCs on HCC development. Interestingly, subcutaneous lesions of the co-transplantation group were more functional in synthesizing and storing glycogen. Collagen and α-SMA cells were also at higher levels in the co-transplantation group than those in the control group. Most importantly, co-transplantation of HepG2 cells with E. granulosus s.l. PSCs led to significant increase in the expression of T cell markers, PD1/PD-L1 and NK cells markers. E. granulosus s.l. may have promoting effects on HCC development, which was closely associated with the immune responses of T cells and NK cells.
免疫细胞是针对寄生虫感染和恶性肿瘤的免疫反应中的关键参与者。大量证据表明，细粒棘球蚴亚种（E. granulosus s.l.）感染和肝细胞癌（HCC）发展之间可能存在可能的关系。因此，本研究旨在观察免疫细胞在移植带有或不带有细粒棘球蚴亚种（E. granulosus s.l.）原头蚴（PSCs）的 HepG2 细胞系后形成皮下病变中的关键作用。在对照组中，将 HepG2 细胞系皮下注射到裸鼠中。在共移植组中，将 HepG2 细胞系与高剂量的细粒棘球蚴亚种（E. granulosus s.l.）PSCs 皮下共注射。从移植后的第 25 天开始，每四天测量一次皮下病变的体积，第 37 天取出进行进一步研究。观察基本的病理和功能变化。此外，通过免疫组织化学染色和实时定量聚合酶链反应（qRT-PCR）分析进一步检测 Ki67、Bcl-2、Caspase3、α-平滑肌肌动蛋白（α-SMA）、T 细胞标志物（CD3、CD4、CD8）、PD1/PD-L1、自然杀伤（NK）细胞标志物（CD16、CD56）的表达。皮下病变的体积逐渐增加，出现了病理异质性的肿瘤细胞，在共移植组中更为明显。与对照组相比，共移植组增殖标志物 Ki67 和 Bcl-2 的表达水平较高。相反，凋亡标志物 Caspase3 在对照组中被高度检测到，表明细粒棘球蚴亚种（E. granulosus s.l.）PSCs 对 HCC 发展有促进作用。有趣的是，共移植组的皮下病变在合成和储存糖原方面的功能更为强大。共移植组的胶原和α-SMA 细胞也高于对照组。最重要的是，将 HepG2 细胞与细粒棘球蚴亚种（E. granulosus s.l.）PSCs 共移植导致 T 细胞标志物、PD1/PD-L1 和 NK 细胞标志物的表达显著增加。细粒棘球蚴亚种（E. granulosus s.l.）可能对 HCC 发展有促进作用，这与 T 细胞和 NK 细胞的免疫反应密切相关。
