Department of Pediatrics, The First Affiliated Hospital of Guangxi Medical University, Nanning, Guangxi, China (mainland).
Department of Hematology, The First Affiliated Hospital of Guangxi Medical University, Nanning, Guangxi, China (mainland).
Med Sci Monit. 2021 Mar 25;27:e929431. doi: 10.12659/MSM.929431.
BACKGROUND Enzymatically inactive chitinase-like protein CHI3L1 is overexpressed in diffuse large B cell lymphoma (DLBCL) patients with PD-L1 imbalance and promotes tumor progression in the microenvironment. Based on this, we investigated how CHI3L1 acts on the proliferation and apoptosis of DLBCL and whether there is a synergy of CHI3L1 in combination with anti-PD-L1 antibodies in vivo. MATERIAL AND METHODS CHI3L1 was detected by quantitative real-time PCR (RT-PCR) and western blot (WB) in B-lymphoma cell lines. CHI3L1 interference plasmids were constructed, and the levels of proliferation, cell cycle, apoptosis, and cell survival were examined in vitro in B-lymphoma cell lines and in vivo in a murine xenograft model by RT-PCR, WB, CCK-8, and flow cytometry. RESULTS CHI3L1 was significantly expressed in SU-DHL-4 cells. CHI3L1-interfered RNA ShRNA-CHI3L1-1 was chosen to be used in the next experiment because it had a better interference effect. Dampened cell proliferation level, arrested cell cycle, reduced protein expressions of cyclin D1 and cyclin D2, and promoted cell apoptosis level were observed after SU-DHL-4 was transfected with ShRNA-CHI3L1-1. Furthermore, we also noticed increased expression of Bcl-2, decreased expressions of bax, cleaved caspase 3 and cleaved PARP, promoted cell survival-related protein p53, and reduced survivin. CONCLUSIONS This study demonstrated that knockdown of CHI3L1 inhibits cancer cell proliferation by regulating cell cycles, promotes cancer cell apoptosis, and enhances the pro-apoptotic effect of anti-PD-L1 antibody both in vivo and in vitro in DLBCL.
在 PD-L1 失衡的弥漫性大 B 细胞淋巴瘤(DLBCL)患者中,酶失活的几丁质酶样蛋白 CHI3L1 过表达,并在微环境中促进肿瘤进展。基于此,我们研究了 CHI3L1 如何作用于 DLBCL 的增殖和凋亡,以及 CHI3L1 是否与抗 PD-L1 抗体在体内具有协同作用。
通过实时定量 PCR(RT-PCR)和蛋白质印迹(WB)检测 B 淋巴瘤细胞系中的 CHI3L1。构建 CHI3L1 干扰质粒,并通过 RT-PCR、WB、CCK-8 和流式细胞术在体外的 B 淋巴瘤细胞系和体内的小鼠异种移植模型中检测增殖、细胞周期、凋亡和细胞存活。
CHI3L1 在 SU-DHL-4 细胞中明显表达。选择 CHI3L1 干扰 RNA ShRNA-CHI3L1-1 用于下一步实验,因为它具有更好的干扰效果。转染 ShRNA-CHI3L1-1 后,SU-DHL-4 细胞的增殖水平降低,细胞周期停滞,cyclin D1 和 cyclin D2 蛋白表达减少,细胞凋亡水平升高。此外,我们还注意到 Bcl-2 表达增加,bax、cleaved caspase 3 和 cleaved PARP 表达减少,促进了与细胞存活相关的蛋白 p53,降低了 survivin。
本研究表明,在 DLBCL 中,敲低 CHI3L1 通过调节细胞周期抑制癌细胞增殖,促进癌细胞凋亡,并增强抗 PD-L1 抗体的促凋亡作用,无论是在体内还是体外。