Gong Yi, Zhang Xi, Chen Rui, Wei Yan, Zou Zhongmin, Chen Xinghua
Department of Hematology, Xinqiao Hospital, The Third Military Medical University, Chongqing, China.
Department of Hematology-oncology, Chongqing Cancer Institute/Hospital, Chongqing, China.
PeerJ. 2017 Jun 13;5:e3457. doi: 10.7717/peerj.3457. eCollection 2017.
To investigate the association of C-MYC protein expression and risk stratification in mantle cell lymphoma (MCL), and to evaluate the utility of C-MYC protein as a prognostic biomarker in clinical practice.
We conducted immunohistochemical staining of C-MYC, Programmed cell death ligand 1 (PD-L1), CD8, Ki-67, p53 and SRY (sex determining region Y) -11 (SOX11) to investigate their expression in 64 patients with MCL. The staining results and other clinical data were evaluated for their roles in risk stratification of MCL cases using ANOVA, Chi-square, and Spearman's Rank correlation coefficient analysis.
Immunohistochemical staining in our study indicated that SOX11, Ki-67 and p53 presented nuclear positivity of tumor cells, CD8 showed membrane positivity in infiltrating T lymphocytes while PD-L1 showed membrane and cytoplasmic positivity mainly in macrophage cells and little in tumor cells. We observed positive staining of C-MYC either in the nucleus or cytoplasm or in both subcellular locations. There were significant differences in cytoplasmic C-MYC expression, Ki-67 proliferative index of tumor cells, and CD8 positive tumor infiltrating lymphocytes (CD8+TIL) among three risk groups ( = 0.000, = 0.037 and =0.020, respectively). However, no significant differences existed in the expression of nuclear C-MYC, SOX11, p53, and PD-L1 in MCL patients with low-, intermediate-, and high risks. In addition, patient age and serum LDH level were also significantly different among 3 groups of patients ( = 0.006 and = 0.000, respectively). Spearman's rank correlation coefficient analysis indicated that cytoplasmic C-MYC expression, Ki-67 index, age, WBC, as well as LDH level had significantly positive correlations with risk stratification ( = 0.000, 0.015, 0.000, 0.029 and 0.000, respectively), while CD8+TIL in tumor microenvironment negatively correlated with risk stratification of patients ( = 0.006). Patients with increased positive cytoplasmic expression of C-MYC protein and decreased CD8+TIL appeared to be associated with a poor response to chemotherapy, but the correlation was not statistically significant.
Our study suggested that assessment of cytoplasmic C-MYC overexpression and cytotoxic T lymphocytes (CTLs) by immunohistochemical staining might be helpful for MCL risk stratification and outcome prediction. However, large cohort studies of MCL patients with complete follow up are needed to validate our speculation.
探讨套细胞淋巴瘤（MCL）中C-MYC蛋白表达与风险分层的关系，并评估C-MYC蛋白作为临床预后生物标志物的效用。
我们对64例MCL患者进行了C-MYC、程序性细胞死亡配体1（PD-L1）、CD8、Ki-67、p53和性别决定区Y（SRY）-11（SOX11）的免疫组织化学染色，以研究它们的表达情况。使用方差分析、卡方检验和Spearman等级相关系数分析评估染色结果和其他临床数据在MCL病例风险分层中的作用。
我们的研究免疫组织化学染色表明，SOX11、Ki-67和p53在肿瘤细胞中呈核阳性，CD8在浸润性T淋巴细胞中呈膜阳性，而PD-L1主要在巨噬细胞中呈膜和胞质阳性，在肿瘤细胞中较少。我们观察到C-MYC在细胞核或细胞质或两个亚细胞位置均呈阳性染色。三个风险组之间在细胞质C-MYC表达、肿瘤细胞的Ki-67增殖指数和CD8阳性肿瘤浸润淋巴细胞（CD8+TIL）方面存在显著差异（分别为P = 0.000、P = 0.037和P = 0.020）。然而，低、中、高风险的MCL患者在核C-MYC、SOX11、p53和PD-L1的表达上没有显著差异。此外，三组患者的年龄和血清乳酸脱氢酶（LDH）水平也有显著差异（分别为P = 0.006和P = 0.000）。Spearman等级相关系数分析表明，细胞质C-MYC表达、Ki-67指数、年龄、白细胞以及LDH水平与风险分层呈显著正相关（分别为P = 0.000、0.015、0.000、0.029和0.000），而肿瘤微环境中的CD8+TIL与患者的风险分层呈负相关（P = 0.006）。C-MYC蛋白细胞质阳性表达增加而CD8+TIL减少的患者似乎与化疗反应不佳有关，但相关性无统计学意义。
我们的研究表明，通过免疫组织化学染色评估细胞质C-MYC过表达和细胞毒性T淋巴细胞（CTLs）可能有助于MCL的风险分层和预后预测。然而，需要对MCL患者进行完整随访且样本量更大的队列研究来验证我们的推测。
