MTCL1在肝癌中表达及其对术后复发的影响

doi:10.3877/cma.j.issn.2095-3232.2021.06.019

中华肝脏外科手术学电子杂志 ›› 2021, Vol. 10 ›› Issue (06) : 622 -628. doi: 10.3877/cma.j.issn.2095-3232.2021.06.019

基础研究

MTCL1在肝癌中表达及其对术后复发的影响

林国桢¹, 代天星², 邓铭彬², 汪国营³^,^†(

)

^1. 518112　深圳市第三人民医院肝脏外科暨肝移植中心；510630　广州，中山大学附属第三医院肝脏外科暨肝移植中心
^2. 510630　广州，中山大学附属第三医院肝脏外科暨肝移植中心
^3. 510120　广州医科大学附属第一医院肝胆外科

收稿日期:2021-08-30 出版日期:2021-09-23
通信作者: 汪国营
基金资助:
十三五科技重大专项(2017ZX10203205-006-001); 国家自然科学基金(81870449，81770648，81670601); 广东省科技计划项目(2017B020209004); 广州市科技计划项目(2014Y2-00544，2014Y2-00200); 广州市健康医疗协同创新项目(201604020001)

Expression of MTCL1 in hepatocellular carcinoma and its effect on postoperative recurrence

Guozhen Lin¹, Tianxing Dai², Mingbin Deng², Guoying Wang³^,^†()

^1. Department of Hepatobiliary Surgery & Liver Transplantation Center, the Third People's Hospital of Shenzhen, Shenzhen 518112, China; Department of Hepatobiliary Surgery and Liver Transplantation Center, the Third Affiliated Hospital of Sun Yat-sen University, Guangzhou 510630, China
^2. Department of Hepatobiliary Surgery and Liver Transplantation Center, the Third Affiliated Hospital of Sun Yat-sen University, Guangzhou 510630, China
^3. Department of Hepatobiliary Surgery, the First Affiliated Hospital of Guangzhou Medical University, Guangzhou 510120, China

Received:2021-08-30 Published:2021-09-23
Corresponding author: Guoying Wang

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引用本文：

林国桢, 代天星, 邓铭彬, 汪国营. MTCL1在肝癌中表达及其对术后复发的影响[J/OL]. 中华肝脏外科手术学电子杂志, 2021, 10(06): 622-628.

Guozhen Lin, Tianxing Dai, Mingbin Deng, Guoying Wang. Expression of MTCL1 in hepatocellular carcinoma and its effect on postoperative recurrence[J/OL]. Chinese Journal of Hepatic Surgery(Electronic Edition), 2021, 10(06): 622-628.

目的

探讨微管交联因子1（MTCL1）在肝细胞癌（肝癌）中表达及其对术后复发的影响。

方法

回顾性分析2015年10月至2018年12月在中山大学附属第三医院行肝癌切除术的76例患者临床病理资料。其中男66例，女10例；年龄27~68岁，中位年龄51岁。患者均签署知情同意书，符合医学伦理学规定。采用实时定量PCR（QRT-PCR）、Western blot和免疫组化法检测肝癌组织MTCL1 mRNA表达，分析MTCL1 mRNA表达与患者生存预后的关系，并采用TCGA数据库验证。采用细胞划痕实验体外研究MTCL1过表达及siRNA沉默MTCL1后肝癌细胞的迁移能力。两组MTCL1相对表达量比较采用t检验。生存分析采用Kaplan-Meier法和Log-rank检验。

结果

肝癌组织MTCL1 mRNA平均相对表达量为3.47±0.86，明显高于癌旁组织的1.38±0.25（t=2.36，P<0.05）。MTCL1蛋白相对表达量为1.07±0.18，明显高于癌旁组织的0.64±0.04（t=2.64，P<0.05）。免疫组化法检测显示肝癌组织中的MTCL1蛋白高表达。MTCL1 mRNA高、低表达组术后3年无瘤生存率分别为24.7%、52.7%，低表达组无瘤生存明显优于高表达组（χ²= 9.28，P<0.05）。TCGA数据库分析显示高表达组术后总体生存时间较低表达组短（HR=1.500，P<0.05）。细胞划痕实验结果显示，MTCL1过表达组97L肝癌细胞伤口愈合率为（7.79±0.31）%，明显高于空载对照组的（3.21±0.41）%（t=8.89，P<0.05）。MTCL1-siRNA组的Hep-G2肝癌细胞伤口愈合率为（9.12±0.75）%，明显低于阴性对照组的（13.79±0.23）%（t=-5.96，P<0.05）。

结论

肝癌组织MTCL1高表达，MTCL1可促进肝癌细胞迁移，高表达患者肝癌切除术后复发率高，预后差。

关键词: 微管交联因子1, 癌，肝细胞, 细胞迁移, 预后

Objective

To investigate the expression level of microtubule cross-linking factor 1(MTCL1) in hepatocellular carcinoma (HCC) and its effect on the postoperative recurrence of HCC.

Methods

Clinicopathological data of 76 HCC patients who underwent hepatectomy in the Third Affiliated Hospital of Sun Yat-sen University from October 2015 to December 2018 were retrospectively analyzed. Among them, 66 patients were male and 10 female, aged from 27 to 68 years, with a median age of 51 years. The informed consents of all patients were obtained and the local ethical committee approval was received. The expression of MTCL1 mRNA in HCC tissues were detected by quantitative real-time PCR (QRT-PCR), Western blot and immunohistochemical staining. The relationship between MTCL1 mRNA expression and survival of patients was analyzed, and was verified by TCGA database. The migration capability of HCC cells with MTCL1 overexpressing and siRNA-silenced MTCL1 was evaluated by cell scratch test in vitro. The relative expression level of MTCL1 between two groups was compared by t test. Kaplan-Meier method andLog-rank test were performed for survival analysis.

Results

The average relative expression of MTCL1 mRNA in HCC tissues was 3.47±0.86, significantly higher than 1.38±0.25 in the adjacent tissues (t=2.36, P<0.05). The relative expression of MTCL1 protein in HCC tissues was 1.07±0.18, significantly higher than 0.64±0.04 in the adjacent tissues (t=2.64, P<0.05). Immunohistochemical staining detected that MTCL1 protein was highly expressed in HCC tissues. The 3-year tumor-free survival rate was 24.7% in MTCL1 mRNA high expression group, and 52.7% in low expression group, the 3-year tumor-free survival in low MTCL1 mRNA expression group was significantly better than that in high expression group (χ²= 9.28, P<0.05). TCGA database analysis demonstrated that the overall survival in the MTCL1 mRNA high expression group was significantly shorter than that in the low expression group (HR=1.500, P<0.05). Cell scratch test showed that the wound healing rate of MHCC97-L cells in MTCL1 overexpression group was (7.79±0.31)%, significantly higher than (3.21±0.41)% in the empty control group (t=8.89, P<0.05). The wound healing rate of Hep-G2 cells in the MTCL1-siRNA group was (9.12±0.75)%, significantly lower than (13.79±0.23)% in the negative control group (t=-5.96, P<0.05).

Conclusions

MTCL1 is highly expressed in HCC tissues, which can promote the migration of HCC cells. Patients with high expression of MTCL1 likely has high recurrence and poor prognosis.

Key words: Microtubule cross-linking factor 1, Carcinoma, hepatocellular, Cell migration, Prognosis

图1 Western blot检测MTCL1蛋白在肝癌组织和癌旁组织中的表达注：MTCL1为微管交联因子1，T为肝癌组织，N为癌旁组织

图2 免疫组化法检测MTCL1蛋白在肝癌组织中的表达注：示MTCL1蛋白在肝癌组织中高表达；MTCL1为微管交联因子1

表1 MTCL1 mRNA表达与肝癌切除术患者临床病理特征及复发的关系（例）

参数		MTCL1 mRNA		χ²值	P值
参数		低表达	高表达	χ²值	P值
性别				0.451	0.502
	男	35	18
	女	2	3
年龄				1.118	0.290
	<60岁	32	15
	≥60岁	5	6
AFP				2.312	0.128
	<400 μg/L	20	7
	≥400 μg/L	17	14
肿瘤直径				2.449	0.118
	<5 cm	22	8
	≥5 cm	15	13
肿瘤数目				0.081	0.776
	单发	26	14
	多发	11	7
BCLC分期				4.337	0.037
	A+B期	34	14
	C期	3	7
肝癌分化程度				1.045	0.307
	高中分化	33	21
	低分化	4	0
血管侵犯				6.009	0.014
	无	34	13
	有	3	8
门静脉癌栓				6.186	0.013
	无	36	15
	有	1	6
复发				5.688	0.017
	否	24	6
	是	13	15

表1 MTCL1 mRNA表达与肝癌切除术患者临床病理特征及复发的关系（例）

参数		MTCL1 mRNA		χ²值	P值
参数		低表达	高表达	χ²值	P值
性别				0.451	0.502
	男	35	18
	女	2	3
年龄				1.118	0.290
	<60岁	32	15
	≥60岁	5	6
AFP				2.312	0.128
	<400 μg/L	20	7
	≥400 μg/L	17	14
肿瘤直径				2.449	0.118
	<5 cm	22	8
	≥5 cm	15	13
肿瘤数目				0.081	0.776
	单发	26	14
	多发	11	7
BCLC分期				4.337	0.037
	A+B期	34	14
	C期	3	7
肝癌分化程度				1.045	0.307
	高中分化	33	21
	低分化	4	0
血管侵犯				6.009	0.014
	无	34	13
	有	3	8
门静脉癌栓				6.186	0.013
	无	36	15
	有	1	6
复发				5.688	0.017
	否	24	6
	是	13	15

图3 MTCL1 mRNA高表达和低表达患者Kaplan-Meier生存曲线注：MTCL1为微管交联因子1

图4 97L肝癌细胞划痕实验注：MTCL1为微管交联因子1，Vector为空载对照组，OE-MTCL1为MTCL1过表达组

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