Bo Jian, Wang Xiaojuan, Li Jie, Wang Wenqing, Zhang Jinqian
Department of Hematology, Chinese People's Liberation Army General Hospital, Beijing 100853, P.R. China.
Department of Hematology, The Second Hospital of Shanxi Medical University, Taiyuan, Shanxi 030001, P.R. China.
Oncol Lett. 2017 Jul;14(1):433-440. doi: 10.3892/ol.2017.6114. Epub 2017 May 3.
Patients with chronic HBV infection have been reported to suffer a significantly increased risk of NHL, but the underlying mechanisms remain to be clearly explained. The aim of the present study was to clarify the relationship between chronic HBV infection and NHL development. Fluorescence-activated cell sorting, Annexin V/7-aminoactinomycin D staining and MTS assay were used to analyze the rate of apoptosis and cell viability. In addition, western blotting was used to detect protein expression. The effects of the activator of SIRT1, SRT1720, and the inhibitor of SIRT1, nicotinamide, were also analyzed. The expression levels cytokines and chemokines were determined by multiplex assay. Hepatitis B surface antigen (HBsAg) was demonstrated to increase the viability of the human peripheral B lymphoblastoid cell line, IM-9, in a dose- and time-dependent manner. HBsAg also decreased histone H3 acetylation and p21 expression at the molecular level. HBsAg upregulated the expression of anti-apoptotic B-cell lymphoma-extra-large and B-cell lymphoma 2 proteins, and inactivated the intrinsic apoptosis pathway by reducing BCL2 associated X, apoptosis regulator expression and increasing the expression of sirtuin 1 (SIRT1) and nuclear factor-κB (NF-κB). HBsAg also altered the levels of certain chemokines and cytokines, including interleukin (IL)-4, -10 and -12, C-X-C motif chemokine 10 and C-C motif chemokine ligand 5. Inhibition of SIRT1 suppressed the effects induced by HBsAg. The anti-apoptotic effect of HBsAg in IM-9 cell lines occurred via the promotion of cell viability, inhibition of apoptosis, regulation of chemokines and cytokines, acetylation of histone H3 and alteration of SIRT1 and NF-κB expression. In conclusion, chronic stimulation with HBsAg promoted the viability of the human B lymphoblastoid cell line, IM-9, through regulation of the SIRT1-NF-κB pathway. This may be an underlying mechanism of HBV-associated NHL.
据报道,慢性乙肝病毒(HBV)感染者患非霍奇金淋巴瘤(NHL)的风险显著增加,但其潜在机制仍有待明确解释。本研究的目的是阐明慢性HBV感染与NHL发生之间的关系。采用荧光激活细胞分选、膜联蛋白V/7-氨基放线菌素D染色和MTS法分析细胞凋亡率和细胞活力。此外,采用蛋白质印迹法检测蛋白质表达。还分析了SIRT1激活剂SRT1720和SIRT1抑制剂烟酰胺的作用。通过多重检测法测定细胞因子和趋化因子的表达水平。结果表明,乙肝表面抗原(HBsAg)以剂量和时间依赖性方式提高人外周B淋巴母细胞系IM-9的活力。HBsAg在分子水平上还降低了组蛋白H3乙酰化和p21表达。HBsAg上调抗凋亡蛋白B细胞淋巴瘤-特大蛋白和B细胞淋巴瘤2蛋白的表达,并通过降低BCL2相关X蛋白(凋亡调节因子)的表达以及增加沉默调节蛋白1(SIRT1)和核因子κB(NF-κB)的表达来使内源性凋亡途径失活。HBsAg还改变了某些趋化因子和细胞因子的水平,包括白细胞介素(IL)-4、-10和-12、C-X-C基序趋化因子10和C-C基序趋化因子配体5。抑制SIRT1可抑制HBsAg诱导的效应。HBsAg在IM-9细胞系中的抗凋亡作用是通过促进细胞活力、抑制凋亡、调节趋化因子和细胞因子、组蛋白H3乙酰化以及改变SIRT1和NF-κB表达而发生的。总之,HBsAg的慢性刺激通过调节SIRT1-NF-κB途径促进了人B淋巴母细胞系IM-9的活力。这可能是HBV相关NHL的潜在机制。