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第42卷第10期
·1358 · 南 京 医 科 大 学 学 报 2022年10月
expression in tissue sections from 121 patients with ESCC. Cox proportional survival risk model was enrolled to analyze the survival
risk factors of ESCC patients,and multivariate correlation analysis was used for the analysis of the association of HMGCS1 with the
clinicopathology of ESCC patients. Furthermore,Kaplan ⁃ Meier analysis was employed to analyze the effect of HMGCS1 on the
prognosis of ESCC patients. The migration and invasion ability of ESCC cells were detected by scratch and Transwell. Results:We
identified 45 significantly up⁃regulated and 51 down⁃regulated(including HMGCS1)proteins in the tumor tissues of patients with lung
metastasis,compared with those without metastasis. GEPIA verified that HMGCS1 was down⁃regulated in esophageal cancer. Western
blot and qPCR results further validated that HMGCS1 expression in ESCC cells was significantly lower than that in normal esophageal
epithelium. Immunohistochemical staining of 121 patients with ESCC confirmed that the expression of HMGCS1 in ESCC was
significantly lower than that in normal esophageal tissues. Additionally,the expression of HMGCS1 in patients with lymph node
metastasis was significantly lower than that in patients without lymph node metastasis(P < 0.05). Cox survival risk analysis showed
that T stage[HR:2.118(1.020~4.399)],lymph node metastasis[HR:2.127(1.466~5.584)]and low HMGCS1 expression were risk
factors[HR:0.413(0.211~0.807)]for survival of ESCC. Multivariate correlation analysis indicated that HMGCS1 expression was
significantly correlated with lymph node metastasis. Kaplan ⁃ Meier survival analysis demonstrated that patients with low HMGCS1
expression had a poor prognosis. Furthermore,we found that overexpression of HMGCS1 markedly inhibited the migration and invasion
ability of ESCC cells(P < 0.05). Conclusion:HMGCS1 could suppress the metastasis of ESCC cells and its expression was positively
correlated with the prognosis of ESCC patients.
[Key words] ESCC;HMGCS1;prognosis;metastasis
[J Nanjing Med Univ,2022,42(10):1357⁃1363,1458]
食管癌是最常见的消化道恶性肿瘤之一,其发
1 材料和方法
病率位居我国消化道肿瘤的第3位 。食管鳞状细
[1]
胞癌(esophageal squamous cell carcinoma,ESCC)是 1.1 材料
最常见的食管恶性肿瘤组织学类型,该组织类型在 人正常食管上皮细胞HEEC购自北京龙跃生物
发展中国家发生率较高,其中超过一半的病例发生 科技发展有限公司,人食管鳞癌细胞 KYSE150、
在中国 。尽管近年来多种新的药物和治疗方法已 KYSE30 购自武汉普诺赛(Procell)生命科技有限公
[2]
经被应用于 ESCC的临床治疗,但ESCC患者的总体 司。DMEM 培养基(Gibco 公司,美国);胎牛血清
预后情况并没有得到显著改善 。转移和复发是食 (FBS)、氨苄青霉素(Hyclone 公司,美国);过表达质
[3]
管鳞癌致死的主要原因,转移被认为是由多个基因 粒由安徽通用生物设计并生产。SDS⁃PAGE 试剂
和多信号通路控制的复杂过程,目前为止其发生的 (上海生工)。BCA Protein Assay Kit、GAPDH抗体购
机制仍未完全阐明。因此,阐明食管鳞癌细胞转移 自 上 海 碧 云 天 公 司 。 Lipofectamine TM 3000 试 剂
的潜在机制将有助于开发新的有效方法来降低 (Thermo Fisher Scientific 公司,美国);HMGCS1 抗
ESCC 患者的病死率。3⁃羟基⁃3⁃甲基戊二酰辅酶 A 体、二抗(Abcam公司,美国)。
合酶 1(3⁃hydroxy⁃3⁃methylglutaryl⁃CoA synthase 1, 本研究所有临床样本均经南京医科大学附属淮
HMGCS1)是一种代谢酶,可将乙酰辅酶A与乙酰乙 安第一医院伦理委员会审批(批号:YX⁃2021⁃107⁃01),
酰辅酶 A 缩合形成 HMG⁃CoA,HMG⁃CoA 是甲羟戊 纳入研究的组织芯片(tissue microarray,TMA)中
[4]
酸途径中的重要底物 。HMGCS1在人类肿瘤中的 121 例 ESCC 患者术后样本均为 2012 年 12 月—
功能研究较少,尤其在食管鳞癌中的功能仍然未 2013年12月在本院行食管鳞癌手术治疗的患者。
知。本研究通过蛋白质谱和测序,筛选出与食管鳞 1.2 方法
癌转移相关的基因 HMGCS1,然后通过临床样本分 1.2.1 TMA免疫组织化学染色
析该基因对食管鳞癌患者预后的影响,同时通过体 石蜡切片常规脱蜡水化,加 0.01 mol/L 枸橼酸
外实验探讨 HMGCS1 对食管鳞癌细胞侵袭迁移能 缓冲液(pH 6.0),用高压锅进行抗原热修复,加 3%
力的影响,以期为食管鳞癌开发新的潜在治疗靶点 H2O2 避光室温孵育 30 min,PBS 洗涤 3 次。滴加
提供方向。 10%的正常非免疫牛血清,37 ℃孵育 1 h,PBS 洗涤