A study on mechanism of GOLM1 regulating PI3K/AKT/mTOR signaling pathway to promote proliferation, invasion and migration of lung adenocarcinoma cells

Haipeng ZHU; Jun HU; Min JIANG; Ruonan CAI; Junqiao WANG; Li LI

doi:10.19401/j.cnki.1007-3639.2022.03.003

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A study on mechanism of GOLM1 regulating PI3K/AKT/mTOR signaling pathway to promote proliferation, invasion and migration of lung adenocarcinoma cells

Article | 更新时间：2025-12-31

- A study on mechanism of GOLM1 regulating PI3K/AKT/mTOR signaling pathway to promote proliferation, invasion and migration of lung adenocarcinoma cells
- China Oncology Vol. 32, Issue 3, Pages: 207-217(2022)
- 作者机构：
  
  1. 克拉玛依市中心医院血液肿瘤科,新疆克拉玛依 834000
  2. 克拉玛依市中心医院病理科,新疆克拉玛依 834000
- 作者简介：
- 基金信息：
- DOI：10.19401/j.cnki.1007-3639.2022.03.003
  CLC： R734.2
- Received：13 September 2021，
  
  Published：30 March 2022
- 稿件说明：
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Haipeng ZHU, Jun HU, Min JIANG, et al. A study on mechanism of GOLM1 regulating PI3K/AKT/mTOR signaling pathway to promote proliferation, invasion and migration of lung adenocarcinoma cells[J]. China Oncology, 2022, 32(3): 207-217.
DOI：

Haipeng ZHU, Jun HU, Min JIANG, et al. A study on mechanism of GOLM1 regulating PI3K/AKT/mTOR signaling pathway to promote proliferation, invasion and migration of lung adenocarcinoma cells[J]. China Oncology, 2022, 32(3): 207-217. DOI： 10.19401/j.cnki.1007-3639.2022.03.003.

摘要

背景与目的：

高尔基体膜蛋白1（Golgi membrane protein 1

GOLM1）在肺腺癌中发挥促癌作用

但对肺腺癌细胞增殖、侵袭和迁移的影响及其作用机制尚不明确。探究GOLM1对肺腺癌细胞增殖、侵袭和迁移的影响及其作用机制。

方法：

选取2019年4月

#x02014;2021年4月在克拉玛依市中心医院行手术切除的肺腺癌患者的癌组织及相应癌旁组织标本各90例

采用免疫组织化学法检测肺腺癌组织和癌旁组织中GOLM1的表达

并分析肺腺癌组织中GOLM1表达与临床病理学特征的关系。采用蛋白质印迹法（Western blot）检测人肺上皮细胞系BEAS-2B及人肺腺癌H460、A549、PG49、H1299细胞系中GOLM1的表达。取对数生长期的肺腺癌A549细胞

随机分为空白组（细胞未转染）、GOLM1小干扰RNA阴性对照（small interfering RNA negative control

si-NC）组（细胞转染si-NC）、GOLM1小干扰RNA（GOLM1 small interfering RNA

si-GOLM1）组（细胞转染si-GOLM1）、胰岛素样生长因子-1（insulin-like growth factor-1

IGF-1）组[10

#x003bc;mol/L的磷脂酰肌醇3-激酶（phosphatidylinositol 3-kinase

PI3K）/蛋白激酶B（protein kinase B

AKT）/哺乳动物雷帕霉素靶蛋白（mammalian target of rapamycin

mTOR）信号转导通路激活剂IGF-1处理30 min

]

和si-GOLM1+IGF-1组（10

#x003bc;mol/L的IGF-1处理30 min后再转染si-GOLM1）

采用细胞计数试剂盒-8（cell counting kit-8

CCK-8）法检测各组肺腺癌A549细胞增殖

采用划痕实验检测各组肺腺癌A549细胞迁移

采用transwell实验检测各组肺腺癌A549细胞侵袭

采用Western blot检测各组肺腺癌A549细胞中GOLM1、PI3K/AKT/mTOR信号转导通路相关蛋白的表达。BALB/c裸小鼠右侧皮下注射肺腺癌A549细胞建立移植瘤模型

分为空白组、si-NC组、si-GOLM1组、IGF-1组和si-GOLM1+IGF-1组

每组6只

注射6周后处死裸小鼠

收集肿瘤并测量肿瘤的质量和体积。

结果：

免疫组织化学结果显示

肺腺癌组织中GOLM1的表达阳性率显著高于癌旁组织（

0.05）。GOLM1的表达与肿瘤分化程度、淋巴结转移、临床分期显著相关（

0.05）

而与肺腺癌患者性别、年龄、是否吸烟无显著相关性（

0.05）。与BEAS-2B细胞比较

人肺腺癌H460、A549、PG49、H1299细胞中GOLM1的相对表达水平显著升高（

0.05）

且肺腺癌A549细胞中GOLM1的相对表达水平最高

因此选用肺腺癌A549细胞进行后续实验。与空白组、si-NC组比较

si-GOLM1组肺腺癌A549细胞的吸光度（

）值、划痕愈合率、侵袭细胞数目、GOLM1、p-PI3K/PI3K、p-AKT/AKT、p-mTOR/mTOR蛋白的相对表达水平显著降低（

0.05）

IGF-1组肺腺癌A549细胞除GOLM1无显著性变化（

0.05）外

其余对应指标均显著升高（

均

0.05）。与si-GOLM1组比较

si-GOLM1+IGF-1组肺腺癌A549细胞的

值、划痕愈合率、侵袭细胞数目、p-PI3K/PI3K、p-AKT/AKT、p-mTOR/mTOR蛋白的相对表达水平显著升高（

0.05）

GOLM1的相对表达水平差异无统计学意义（

0.05）。与空白组、si-NC组比较

si-GOLM1组移植瘤的质量和体积显著降低

IGF-1组裸鼠移植瘤的质量和体积显著升高（

0.05）。与si-GOLM1组比较

si-GOLM1+IGF-1组移植瘤的质量和体积显著升高（

0.05）。

结论：

沉默

GOLM

1基因可抑制PI3K/AKT/mTOR信号转导通路的激活

从而抑制肺腺癌A549细胞增殖、迁移和侵袭。

Abstract

Background and purpose:

Golgi membrane protein 1 (GOLM1) plays the role of an oncogene in lung adenocarcinoma (LUAD)

however

the effects of GOLM1 on the proliferation

invasion and migration of LUAD cells and its mechanism are not clear yet. This study investigated the effects of GOLM1 on the proliferation

invasion and migration of LUAD cells and its mechanism of action.

Methods:

We selected cancer tissues and corresponding paracancerous tissue specimens from 90 LUAD patients who underwent surgical resection in Karamay Central Hospital from April 2019 to April 2021. The expression of GOLM1 protein in LUAD tissues and paracancerous tissues was detected by immunohistochemistry

and the relationship between GOLM1 protein expression and clinicopathological characteristics of LUAD tissues was analyzed. Western blot was used to det

ect the expression of GOLM1 protein in human lung epithelial cells BEAS-2B and human lung adenocarcinoma H460

A549

PG49 and H1299 cells. Lung adenocarcinoma A549 cells in logarithmic growth stage were randomly divided into blank group (cells not transfected)

GOLM1 small interfering RNA negative control (si-NC) group (cells transfected with si-NC)

GOLM1 small interfering RNA (si-GOLM1) group (cells transfected with si-GOLM1)

insulin-like growth factor-1 (IGF-1) group [10

#x003bc;mol/L phosphatidylinositol 3-kinase (PI3K)/protein kinase B (AKT)/mammalian target of rapamycin (mTOR) signaling pathway activator IGF-1 for 30 min

]

and si-GOLM1+IGF-1 group (after treatment with 10

#x003bc;mol/L IGF-1 for 30 min

si-GOLM1 was transfected). Cell counting kit-8 method was used to detect cell proliferation in each group of lung adenocarcinoma A549 cells. Scratch test was used to detect cell migration in each group of lung adenocarcinoma A549 cells. Transwell experiment was used to detect cell invasion in each group of lung adenocarcinoma A549 cells. Western blot was used to detect the expressions of GOLM1 and PI3K/AKT/mTOR signaling pathway related proteins in each group of lung adenocarcinoma A549 cells. Xenograft model was constructed by subcutaneously injecting A549 cells on the right side of BALB/c nude mice

which were divided into: nude mice blank group

nude mice si-NC group

nude mice si-GOLM1 group

nude mice IGF-1 group

nude mice si-GOLM1+IGF-1 group

with 6 mice in each group. The nude mice were sacrificed six weeks after injection

the tumor was collected

and the weight and volume of the tumor were measured.

Results:

The results of immunohistochemistry showed that the positive expression rate of GOLM1 protein was significantly higher in LUAD tissues than in adjacent tissues (

0.05). The expression of GOLM1 protein was significantly correlated with the degree of tumor differentiation

lymph node metastasis

and clinical stage (

0.05)

but not

significantly correlated with gender

age and smoking status of LUAD patients (

0.05). Compared with BEAS-2B cells

the relative expression level of GOLM1 protein in human lung adenocarcinoma H460

A549

PG49 and H1299 cells was significantly increased (

0.05)

and the relative expression level of GOLM1 protein in A549 cells was the highest. Therefore

A549 cells were selected for subsequent experiments. Compared with the blank group and the si-NC group

OD value

scratch healing rate

number of invaded cells

GOLM1

p-PI3K/PI3K

p-AKT/AKT and p-mTOR/mTOR protein relative expression levels in the lung adenocarcinoma A549 cells of the si-GOLM1 group were significantly reduced (

0.05). In the lung adenocarcinoma A549 cells of the IGF-1 group

there was no significant change in GOLM1 protein

and the other corresponding indicators were significantly increased (

0.05). Compared with the si-GOLM1 group

the OD value

scratch healing rate

number of invaded cells

p-PI3K/PI3K

p-AKT/AKT and p-mTOR/mTOR protein relative expression levels of lung adenocarcinoma A549 cells in the si-GOLM1+IGF-1 group were significantly increased (

0.05)

and there was no significant difference in GOLM1 protein relative expression level (

0.05). Compared with the nude mice blank group and nude mice si-NC group

the mass and volume of transplanted tumors in the nude mice si-GOLM1 group were significantly reduced

while the mass and volume of transplanted tumors in the nude mice IGF-1 group were significantly increased (

0.05). Compared with the nude mice si-GOLM1 group

the mass and volume of transplanted tumors in the nude mice si-GOLM1+IGF-1 group were significantly increased (

0.05).

Conclusion:

Silencing

GOLM

1 gene can inhibit the activation of PI3K/AKT/mTOR signaling pathway

thereby inhibiting the proliferation

migration and invasion of lu

ng adenocarcinoma A549 cells.

关键词

Keywords

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