Role and mechanism of a novel fusion gene RELCH-RET in driving malignant transformation of human bronchial epithelial cells: a preliminary study

ZHOU Xiaogang; HAO Xianglin; XIA Jiying; HUANGFU Zhimin; FU Wanlei; LYU Yangfan; GUO Qiaonan

doi:10.16016/j.2097-0927.202502092

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Role and mechanism of a novel fusion gene RELCH-RET in driving malignant transformation of human bronchial epithelial cells: a preliminary study

Basic Medicine | 更新时间：2025-07-14

- Role and mechanism of a novel fusion gene RELCH-RET in driving malignant transformation of human bronchial epithelial cells: a preliminary study
- Journal of Army Medical University Vol. 47, Issue 13, Pages: 1444-1453(2025)
- 作者机构：
  
  1.陆军军医大学（第三军医大学）第二附属医院病理科，重庆
  2.中国人民解放军中部战区总医院呼吸与危重症医学科，湖北武汉
- 作者简介：
  
  GUO Qiaonan， E-mail： qiaonan85@263.net
- 基金信息：
  
  the National Natural Science Foundation for Yong Scholars of China(82002446)
- DOI：10.16016/j.2097-0927.202502092
  CLC： R394.3;R730.23;R734.1
- Received：03 March 2025，
  
  Revised：12 May 2025，
  
  Published：15 July 2025
- 稿件说明：
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周小刚，郝翔麟，夏吉影，等.新融合基因RELCH-RET驱动人支气管上皮细胞恶性转化的作用及机制的初步探究［J］.陆军军医大学学报，2025，47（13）：1444-1453.DOI：10.16016/j.2097-0927.202502092.

ZHOU X G，HAO X L，XIA J Y，et al.Role and mechanism of a novel fusion gene RELCH-RET in driving malignant transformation of human bronchial epithelial cells： a preliminary study［J］.J Army Med Univ，2025，47（13）：1444-1453.DOI：10.16016/j.2097-0927.202502092.
周小刚，郝翔麟，夏吉影，等.新融合基因RELCH-RET驱动人支气管上皮细胞恶性转化的作用及机制的初步探究［J］.陆军军医大学学报，2025，47（13）：1444-1453.DOI：10.16016/j.2097-0927.202502092. DOI：

ZHOU X G，HAO X L，XIA J Y，et al.Role and mechanism of a novel fusion gene RELCH-RET in driving malignant transformation of human bronchial epithelial cells： a preliminary study［J］.J Army Med Univ，2025，47（13）：1444-1453.DOI：10.16016/j.2097-0927.202502092. DOI：

摘要

目的

探讨新融合基因

RELCH-RET

驱动正常人支气管上皮（human bronchial epithelial，HBE）细胞发生恶性转化的作用及初步机制。

方法

本研究基于2019年1月至2022年6月陆军军医大学第二附属医院非小细胞肺癌（non-small c

ell lung cancer，NSCLC）患者回顾性临床数据（

=469），筛选出

RELCH-RET

融合基因作为研究靶点。为进一步验证其功能及机制，构建3组细胞模型：阴性对照组（HBE VC，转染空载慢病毒）、RET对照组（HBE RET，转染Flag-RET过表达慢病毒）及实验组（HBE RELCH-RET，转染Flag-RELCH-RET过表达慢病毒）；MTS检测细胞增殖情况，Transwell实验分析细胞迁移、侵袭能力；利用15只4周龄雌性肥胖型糖尿病/严重联合免疫缺陷（non-obese diabetic/severe combined immunodeficiency，NOD/SCID）小鼠（SPF级），体质量（15.1±0.4）g，随机分为3组（

=5），通过皮下移植瘤实验评估体内成瘤能力；Western blot检测RET酪氨酸激酶（Y905）自磷酸化水平及其下游信号通路关键蛋白ERK1/2、EGFR（Y845）和STAT3（Y705）的磷酸化水平；采用化学交联法结合Western blot分析RELCH-RET的二聚化及多聚化状态，并通过解交联实验验证其可逆性。

结果

在NSCLC患者中筛选出3例阳性

RELCH-RET

融合基因患者；与HBE VC及HBE RET组相比，HBE RELCH-RET组细胞增殖能力显著增强（

0.01），且获得迁移及侵袭能力（

0.01），而对照组未表现出迁移及侵袭能力；小鼠异种移植瘤实验可见HBE RELCH-RET组形成明显肿瘤结节（

0.001），而对照组无肿瘤生长；Western blot结果提示，

RELCH-RE

T可诱导RET酪氨酸残基（Y905）发生磷酸化自激活，并显著上调ERK1/2、EGFR（Y845）及STAT3（Y705）蛋白的磷酸化水平；化学交联-Western blot分析证实RELCH-RET在HBE细胞中形成分子量约为170 kDa的二聚体，经解交联处理后可恢复至单体状态。

结论

本研究初步揭示了RELCH-RET通过以不依赖配体方式二聚化或多聚化介导RET磷酸化自激活，进而激活下游典型RET信号通路，最终驱动正常HBE细胞恶性转化。

Abstract

‍Objective‍　‍To investigate the role and primary mechanism of a novel fusion gene

RELCH-RET

in driving the malignant transformation of normal human bronchial epithelial (HBE) cells. Methods‍　‍Based on retrospective clinical data from 456 non-small cell lung cancer (NSCLC) patients admitted in the Second Affiliated Hospital of Army Medical University from January 2019 to June 2022

a fusion gene

RELCH-RET

was identified as a research target. Three cell models were established: negative control (HBE VC

transfected with empty lentiviral vector)

RET control (HBE RET

transfected with

lentiviral overexpression vector of Flag-RET)

and experimental group (HBE RELCH-RET

transfected with lentiviral overexpression vector of Flag-RELCH-RET). MTS assay and Transwell

assay were used to detect cell proliferation and migratory and invasive abilities.

In vivo

tumorigenicity of the 3 cell models was assessed in 15 female non-obese diabetic/severe combined immunodeficiency (NOD/SCID) mice (SPF grade

4 weeks old

weighing 15.1±0.4 g)

via

subcutaneous xenograft experiments

with 5 animals in each group. Western blotting was employed to detect the autophosphorylation of RET (Y905) and the phosphorylation of downstream signaling proteins ERK1/2

EGFR (Y845) and STAT3 (Y705). Dimerization and multimerization status of RELCH-RET were analyzed by chemical cross-linking (DTME treatment) in combination with Western blotting

with the reversibility being confirmed through de-cross-linking experiments. Results‍　‍There were 3 cases carrying

RELCH-RET

fusion gene screened out from the 469 NSCLC patients. Compared with the HBE VC and HBE RET groups

the HBE RELCH-RET group exhibited significantly enhanced cell proliferation (

0.01)

and acquired migratory and invasive abilities (

0.01)

while the control groups did not demonstrate the abilities. In the mouse xenograft tumor model

HBE cells stably expressing RELCH-RET developed significant tumor nodules (

0.001)

whereas the control groups (empty vector and wild-type RET) failed to exhibit detectable tumor growth. Western blotting revealed that

RELCH-RET

could induce the autophosphorylation of the RET tyrosine residue (Y905) and significantly up-regulate the phosphorylation levels of ERK1/2

EGFR (Y845)

and STAT3 (Y705) proteins. Chemical cross-linking combined with Western blot analysis demonstrated that RELCH-RET formed a dimer (~170 kDa) in HBE cells

which is reversibly dissociated into monomers upon decross-linking treatment. Conclusion‍　‍The novel fusion gene

RELCH-RET

promotes ligand-independent dimerization/oligomerization

thereby mediating RET autophosphorylation

subsequently activate

s the downstream typical RET signaling pathway and ultimately drives the malignant transformation of normal HBE cells.

关键词

Keywords

references

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