Cheng Yong-Feng, Shen Yi-Ping, Wang Xue-Mei, Li Dan-Lu, Fan Chun-Yan, Maimaiti Gulibaha, Yan Mei
Department of Pediatrics, First Affiliated Hospital of Xinjiang Medical University, Urumqi 830054, China.
Zhongguo Dang Dai Er Ke Za Zhi. 2024 Nov 15;26(11):1218-1224. doi: 10.7499/j.issn.1008-8830.2404153.
To investigate the mechanism by which mycobacterial antigen 85B (Ag85B) inhibits autophagy and promotes apoptosis in Hodgkin lymphoma (HL) cells.
The clinical data and pathological tissue slides were retrospectively collected from 80 HL children and 30 children with reactive lymphadenopathy (control group) treated at the First Affiliated Hospital of Xinjiang Medical University. Immunohistochemical analysis was performed to assess the expression of microtubule-associated protein 1 light chain 3 (LC3), sequestosome 1 (P62/SQSTM1), and Beclin-1 in the pathological tissues of HL and control groups. Human Hodgkin lymphoma cells (HDLM-2) were divided into the HDLM-2 group and the HDLM-2+Ag85B groups (with Ag85B concentrations of 0.5, 1, 2, and 4 μg/mL). The CCK8 method was used to measure HDLM-2 cell proliferation; qRT-PCR was employed to detect the expression of LC3, P62, Beclin-1, Akt, and mTOR mRNA in cells. An apoptosis kit was used to detect cell apoptosis.
The positive expression of LC3 and Beclin-1 in the HL group were higher than those in the control group (<0.05), while the positive expression of P62 was lower than that in the control group (<0.05). In stages III-IV compared to stages I-II, the positive expression of LC3 and Beclin-1 increased, while the positive expression of P62 decreased (<0.05). Cell experiment results showed that the HDLM-2+Ag85B group had suppressed cell proliferation compared to the HDLM-2 group, with decreased mRNA expression of LC3 and Beclin-1, and increased mRNA expression of P62, PI3K, Akt, and mTOR, leading to increased cell apoptosis. Notably, when Ag85B was at a concentration of 2 μg/mL, it had the strongest effect on HDLM-2 cells after 24 hours (<0.05).
Autophagy is enhanced in children with HL and increases with disease stage. Ag85B can inhibit the proliferation and autophagy of HL tumor cells and promote apoptosis, possibly related to the activation of the PI3K/Akt/mTOR pathway.
探讨分枝杆菌抗原85B（Ag85B）抑制霍奇金淋巴瘤（HL）细胞自噬并促进其凋亡的机制。
回顾性收集新疆医科大学第一附属医院收治的80例HL患儿及30例反应性淋巴结病患儿（对照组）的临床资料及病理组织切片。采用免疫组织化学分析法评估HL组和对照组病理组织中微管相关蛋白1轻链3（LC3）、隔离小体1（P62/SQSTM1）和Beclin-1的表达。将人霍奇金淋巴瘤细胞（HDLM-2）分为HDLM-2组和HDLM-2+Ag85B组（Ag85B浓度分别为0.5、1、2和4μg/mL）。采用CCK8法检测HDLM-2细胞增殖；采用qRT-PCR检测细胞中LC3、P62、Beclin-1、Akt和mTOR mRNA的表达。使用凋亡试剂盒检测细胞凋亡。
HL组中LC3和Beclin-1的阳性表达高于对照组（<0.05），而P62的阳性表达低于对照组（<0.05）。与Ⅰ-Ⅱ期相比，Ⅲ-Ⅳ期LC3和Beclin-1的阳性表达增加，而P62的阳性表达降低（<0.05）。细胞实验结果显示，与HDLM-2组相比，HDLM-2+Ag85B组细胞增殖受到抑制，LC3和Beclin-1的mRNA表达降低，P62、PI3K、Akt和mTOR的mRNA表达增加，导致细胞凋亡增加。值得注意的是，当Ag85B浓度为2μg/mL时，24小时后对HDLM-2细胞的作用最强（<0.05）。
HL患儿的自噬增强，并随疾病分期增加。Ag85B可抑制HL肿瘤细胞的增殖和自噬并促进凋亡, 可能与PI3K/Akt/mTOR通路的激活有关。
