|
摘要：
|
|
摘要：目的 分析微小核糖核酸（microRNA，miR）-16与肝细胞癌临床病理特征、预后的关系，并探讨
其对肝癌细胞侵袭转移的影响。方法 选取2014年1月至2016年1月于北京市顺义区医院诊治的88例肝细
胞癌患者为研究对象，采用实时荧光定量聚合酶链反应（quantitative real-time polymerase chain reaction，
qRT-PCR）检测患者肝癌组织和癌旁正常组织中miR-16的相对表达水平，以肝癌组织miR-16相对表达
量的平均数为分界点，大于或等于该值者为miR-16高表达组，低于该值者为miR-16低表达组，并分
析两组患者肿瘤直径、Edmondson分级、包膜受侵、门脉受侵及肝内转移情况及TNM分期的差异。对
所有研究对象随访3年，记录患者无进展生存期（progression-free-survival，PFS）和总生存期（overall
survival，OS），采用Kaplan-Meier法绘制生存曲线并分析miR-16低表达组和miR-16高表达组生存率的
差异，用Cox回归模型探讨影响肝细胞癌预后的独立危险因素。采用miR-16过表达质粒和空载质粒转染
肝癌细胞系Huh-7，建立miR-16过表达组和阴性对照组，同时设立空白对照组（未给予干预措施）；用
Transwell实验检测miR-16在体外肝癌细胞侵袭转移中的作用。结果 miR-16在肝癌组织和癌旁正常组织
中相对表达量分别为0.86 ± 0.19和1.85 ± 0.47，差异有统计学意义（t = 4.009，P = 0.016）。miR-16低表达
组包膜受侵、门脉受侵、肝内转移及TNM Ⅲ期发生率分别为76.7%（23/30）、53.3%（16/30）、56.7%
（17/30）、66.7%（20/30）；miR-16高表达组上述指标分别为43.1%（25/58）、43.1%（17/58）、31.0%
（18/58）、37.9%（22/58），差异均有统计学意义（χ 2 值分别为8.954、4.869、5.423、6.544，P值分别
为0.003、0.027、0.020、0.011）。miR-16低表达组肿瘤直径≤ 5 cm比例为40.0%（12/30），Edmondson
Ⅰ～Ⅱ级比例为46.7%（14/30）；miR-16高表达组上述指标分别为55.2%（32/58）、63.8%（37/58），
差异无统计学意义（χ 2 值分别为1.821、2.380，P值分别为0.177、0.123）。随访3年，miR-16低表达组和
高表达组患者PFS分别为（22.90 ± 2.17）个月和（29.47 ± 1.24）个月，差异有统计学意义（t = 18.094，
P ＜ 0.001）；miR-16低表达组和高表达组患者OS分别为（24.87 ± 2.01）个月和（31.00 ± 1.03）个
月，差异有统计学意义（t = 18.966，P ＜ 0.001）。miR-16低表达组和miR-16高表达组无进展生存率
[33.33%（10/30）vs 58.62%（34/58）]和总生存率[40.00%（12/30）vs 65.52%（38/58）]的差异有统计
学意义（χ 2 = 6.888，P = 0.009；χ 2 = 7.046，P = 0.008）。Cox回归分析表明，门脉受侵、肝内转移及
TNM Ⅲ期是影响肝癌预后的独立危险因素（HR = 1.837，95%CI：1.627～2.345；HR = 1.378，95%CI：
1.132～1.534；HR = 1.869，95%CI：1.705～2.432）。空白对照组、阴性对照组和miR-16过表达组穿透生
物膜细胞数分别为（85.67 ± 6.03）个、（86.67 ± 3.51）个和（21.00 ± 3.61）个，组间差异有统计学意义
（F = 63.874，P ＜ 0.001），其中miR-16过表达组显著少于空白对照组和阴性对照组，差异有统计学意义
（t = 15.938，P ＜ 0.001；t = 22.590，P ＜ 0.001），空白对照组和阴性对照组间差异无统计学意义（t =
0.248，P = 0.816）。结论 miR-16在肝癌细胞中低表达，高miR-16相对表达水平可抑制肝癌细胞的侵袭和
转移，有利于患者预后，其中门脉受侵、肝内转移及TNM Ⅲ期是影响肝细胞癌预后的独立危险因素。
|
|
Abstract: Objective To analyze the relationship among microRNA (miR) -16, clinicopathological features
and prognosis of hepatocellular carcinoma, and to explore its influence on invasion and metastasis of
hepatocellular carcinoma cells. Methods Total of 88 patients with primary hepatocellular carcinoma
treated in the Hospital of Shunyi District, Beijing from January 2014 to January 2016 were selected as the
study subjects. Quantitative real-time polymerase chain reaction (qRT-PCR) was used to detect the level
of microRNA-16 in hepatocellular carcinoma tissues and adjacent normal tissues, the average relative
expression of miR-16 in liver cancer tissue was taken as the dividing point, the miR-16 high expression group
of miR-16 was defined as miR-16 ≥ average value, and miR-16 low expression group was defined as miR-16 ＜
average value, The differences of tumor diameter, Edmondson classification, capsule invasion, portal vein
invasion, intrahepatic metastasis and TNM stage between the two groups were analyzed. All subjects were
followed up for 3 years, the progression-free survival (PFS) and overall survival (OS) were recorded. Kaplan-
Meier method was used to draw survival curves and analyze the survival differences between miR-16 high
expression group and miR-16 low expression group. Cox regression model was used to explore independent
risk factors affecting the prognosis of hepatocellular carcinoma. MiR-16 overexpression plasmid and empty
plasmid were transfected into Huh-7 cells to establish the overexpression group and negative control group,
and blank control group was set up at the same time (no intervention was given). Transwell assay was used to
detect the role of microRNA-16 in invasion and metastasis of hepatocellular carcinoma cells in vitro. Results
The relative expression of miR-16 in hepatocellular carcinoma tissues and adjacent normal tissues were 0.86 ± 0.19
and 1.85 ± 0.47, respectively, the difference was statistically significant (t = 4.009, P = 0.016). The capsular
invasion, portal vein invasion, intrahepatic metastasis and TNM phase Ⅲ presence rates of patients in miR-
16 low expression group were 76.7% (23/30), 53.3% (16/30), 56.7% (17/30) and 66.7% (20/30), respectively,
and in miR-16 high expression group were 43.1% (25/58), 43.1% (17/58), 31.0% (18/58) and 37.9% (22/58),
respectively, the differences were statistically significant (χ 2 = 8.954, 4.869, 5.423, 6.544; P = 0.003, 0.027,
0.020, 0.011). The ratio of patients with tumor diameter ≤ 5 cm and Edmondson grade Ⅰ～Ⅱ in miR-16
low expression group were 40.0% (12/30) and 46.7% (14/30), respectively, and in miR-16 high expression
group were 55.2% (32/58) and 63.8% (37/58), respectively, the differences were statistically significant
(χ 2 = 1.821, 2.380; P = 0.177, 0.123). After 3 years of follow-up, the PFS time of patients in miR-16 low
expression group and miR-16 high expression group were (22.90 ± 2.17) months and (29.47 ± 1.24) months,
respectively, the difference was statistically significant (t = 18.094, P ＜ 0.001); the OS time of patients in
miR-16 low expression group and miR-16 high expression group were (24.87 ± 2.01) months and (31.00 ±
1.03) months, respectively, the difference was statistically significant (t = 18.966, P ＜ 0.001). The survival
rates of PFS [33.33% (10/30) vs 58.62% (34/58)] and OS [40.00% (12/30) vs 65.52% (38/58)] of patients in
miR-16 low expression group and miR-16 high expression group were statistically significant (χ 2 = 6.888, P = 0.009;
χ 2 = 7.046, P = 0.008). Cox regression analysis showed that portal vein invasion, intrahepatic metastasis and
TNM phase Ⅲ were independent risk factors affecting the prognosis of hepatocellular carcinoma (HR = 1.837,
95%CI: 1.627～2.345; HR = 1.378, 95%CI: 1.132～1.534; HR = 1.869, 95%CI: 1.705～2.432). The number
of penetrating biofilm cells in blank control group, negative control group and miR-16 over expression group
was 85.67 ± 6.03, 86.67 ± 3.51 and 21.00 ± 3.61, respectively, the difference was statistically significant (F =
63.874, P ＜ 0.001), the number of penetrating biofilm cells in miR-16 over expression group was statistically
lower than that in blank control group and negative control group, respectively (t = 15.938, P ＜ 0.001; t = 22.590,
P ＜ 0.001), there was no significant difference between blank control group and negative control group (t = 0.248,
P = 0.816). Conclusions MiR-16 was lowly expressed in hepatoma cells, the high expression of miR-16 inhibits the
invasion and metastasis of hepatocarcinoma cells, which is beneficial to the prognosis. The invasion of portal vein,
intrahepatic metastasis and TNM stage Ⅲ are independent risk factors for the prognosis of hepatocellular carcinoma.
|
|
基金项目：
|
|
|
|
作者简介：
|
|
|
|
参考文献：
|
|
|
|
服务与反馈：
|
|
【文章下载】【加入收藏】
|
|
|
|