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摘要：
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摘要：目的 探讨微小RNA（microRNA，miRNA）-320a和钠氢交换调控因子1（Na + /H + exchanger
regulatory factor 1，NHERF1）在肝细胞癌（hepatocellular carcinoma，HCC）中的表达及机制。方
法 收集首都医科大学附属北京佑安医院2015年1月至2016年1月经手术治疗的HCC患者肝癌组织及癌
旁组织。采用反转录聚合酶链式反应（reverse transcription-polymerase chain reaction，RT-PCR）检测
miR-320a和NHERFl在HCC组织、癌旁组织、HCC细胞株Bel-7402和正常肝细胞株HL-7702中的表达。
将体外培养的Bel-7402细胞分为对照组、空白组和miR-320a转染组，空白组中仅加入2 ml全培养基；
对照组加入2 ml空载体质粒；miR-320a组将稀释的miR-320a混合液加入到完全养基中，最终体积为
2 ml；采用RT-PCR检测Bel-7402细胞中miR-320a、NHERF1及β-catenin的表达，采用流式细胞术检测
Bel-7402细胞的凋亡，采用Transwell检测Bel-7402细胞迁移侵袭能力。结果 miR-320a（0.51 ± 0.01 vs
0.83 ± 0.02）和NHERFl（0.78 ± 0.02 vs 1.42 ± 0.05）在肝癌组织中的表达均显著低于癌旁组织，差异
有统计学意义（t值分别为-80.25、-68.05，P均＜ 0.001）。miR-320a（0.75 ± 0.03 vs 0.81 ± 0.04）和
NHERFl（0.79 ± 0.05 vs 1.58 ± 0.05）在Bel-7402细胞中的相对表达量均显著低于HL-7702，差异有统
计学意义（t = -2.73，P = 0.021，t = -27.60，P ＜ 0.001）。对照组、空白组和miR-320a转染组在Bel-
7402细胞中miR-320a相对表达量分别为0.77 ± 0.04、0.79 ± 0.05和1.28 ± 0.07，差异有统计学意义（H =
11.66，P = 0.003），miR-320a转染组显著高于对照组和空白组（H值分别为8.308、8.308，P值分别
为0.004、0.004）。对照组、空白组和miR-320a转染组Bel-7402细胞中NHERFl相对表达量分别为0.82 ±
0.04、0.70 ± 0.04和1.46 ± 0.06，差异有统计学意义（H = 15.17，P = 0.001），miR-320a转染组显著
高于对照组和空白组（H值分别为8.337、8.308，P值分别为0.004、0.004）。空白组、对照组和miR-
320a转染组Bel-7402细胞凋亡率分别为11.2%、11.4%、32.5%，差异有统计学意义（χ 2 = 9263.95，
P ＜ 0.001）。其中miR-320a转染组显著高于空白组和对照组（χ 2 值分别为7508.35、5100.96，P均＜
0.001），空白组和对照组间差异无统计学意义（χ 2 = 1.024，P = 0.311）。miR-320a组Bel-7402迁移能
力显著降低。对照组、空白组和miR-320a转染组Bel-7402细胞中β-catenin的相对表达量分别为1.66 ±
0.07、1.62 ± 0.06、0.64 ± 0.02，差异有统计学意义（H = 12.117，P = 0.002）。其中，miR-320a转染
组显著低于空白组和对照组（H值分别为8.308、8.308，P值分别为0.004、0.004），空白组和对照组
间差异无统计学意义（H = 1.641，P = 0.200）。结论 miR-320a和NHERF1在HCC中表达降低，miR-
320a可能通过Wnt信号转导通路发挥作用，抑制癌细胞的增殖。
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Abstract: Objective To investigate the expression and mechanism of microRNA (miRNA)-320a and Na + /
H + exchanger regulatory factor 1 (NHERF1) on hepatocellular carcinoma (HCC). Methods The liver cancer
tissues and adjacent tissues of patients with HCC who were operated on in Beijing YouAn Hospital, Capital
Medical University from January 2015 to January 2016 were collected. Reverse transcription-polymerase
chain reaction (RT-PCR) was used to detect the expression of miR-320a and NHERF1 in HCC tissues,
adjacent tissues, HCC cell line BEL-7402 and normal liver cell line HL-7702. BEL-7402 cells were divided
into control group, blank group and miR-320a transfection group. The blank group was added with 2 ml
medium; the control group was added with 2 ml plasmid; the miR-320a transfection group was added with
diluted miR-320a mixture into the medium, and the final volume was 2 ml. RT-PCR was used to detect
the expression of miR-320a, NHERF1 and β-catenin in Bel-7402 cells, flow cytometry was used to detect
the apoptosis of Bel-7402 cells, and Transwell was used to detect the migration ability of Bel-7402 cells.
Results The expression of miR-320a (0.51 ± 0.01 vs 0.83 ± 0.02) and NHERFl (0.78 ± 0.02 vs 1.42 ± 0.05)
in liver cancer tissues were significantly lower than those in adjacent tissues, the differences were statistically
significant (t = -80.25, -68.05; P ＜ 0.001). The expression of miR-320a (0.75 ± 0.03 vs 0.81 ± 0.04) and
NHERFl (0.79 ± 0.05 vs 1.58 ± 0.05) in Bel-7402 cells were significantly lower than those in HL-7702 cells,
the differences were statistically significant (t = -2.73, P = 0.021; t = -27.60, P ＜ 0.001). The expression of
miR-320a in Bel-7402 cells in control group, blank group and miR-320a transfection group were 0.77 ± 0.04,
0.79 ± 0.05 and 1.28 ± 0.07, the difference was statistically significant (H = 11.66, P = 0.003). The expression
of miR-320a in Bel-7402 cells in miR-320a transfection group was significantly higher than those in control
group and blank group (H = 8.308, 8.308; P = 0.004, 0.004). The expression of NHERFl in Bel-7402 cells in
control group, blank group and miR-320a transfection group were 0.82 ± 0.04, 0.70 ± 0.04 and 1.46 ± 0.06,
the difference was statistically significant (H = 15.17, P = 0.001). The expression of NHERFl in Bel-7402
cells in miR-320a transfection group was significantly higher than those in control group and blank group
(H = 8.337, 8.308; P = 0.004, 0.004). The apoptosis rate of Bel-7402 cells in control group, blank group and
miR-320a transfection group were 11.2%, 11.4% and 32.5%, respectively, the difference was statistically
significant (χ 2 = 9263.95, P ＜ 0.001). The apoptosis rate of Bel-7402 cells in miR-320a transfection group
was significant higher than those in blank group and control group (χ 2 = 7508.35, 5100.96; all P ＜ 0.001).
There was no significant difference between blank group and control group (χ 2 = 1.024, P = 0.311). The
migration ability of BEL-7402 cells reduced significantly. The relative expression of β-catenin in Bel-7402
cells in control group, blank group and miR-320a transfection group were 1.66 ± 0.07, 1.62 ± 0.06 and 0.64 ±
0.02, respectively, the difference was statistically significant (H = 12.117, P = 0.002). The relative expression
of β-catenin in Bel-7402 cells in miR-320a transfection group was significant lower than those in blank group
and control group, (H = 8.308, 8.308; P = 0.004, 0.004). There was no significant difference between blank
group and control group (H = 1.641, P = 0.200). Conclusions The expression of miR-320a and NHERF1
decreased in HCC. MiR-320a may inhibit the proliferation of cancer cells by Wnt signal transduction pathways
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