GPRC5A调控的ABCB1表达对肺腺癌增殖的影响

doi:10.24920/004216

摘要/Abstract

摘要：

目的 ATP结合盒B亚家族成员 1（ATP binding cassette subfamily B member 1，ABCB1）的异常表达在多种癌症的发生发展中发挥关键作用。然而，G蛋白偶联受体C家族5组A型（G protein coupled receptor family C group 5 type A，GPRC5A）调控的ABCB1表达对肺腺癌增殖的影响仍不清楚。本研究探讨了GPRC5A调控的ABCB1表达对肺腺癌增殖的影响。

方法我们采用RT-PCR、Western-blot或免疫组化实验，分析ABCB1在肺腺癌细胞系、人肺腺癌组织以及GPRC5A基因敲除小鼠和野生型小鼠的气管上皮细胞和肺组织中的表达。采用细胞计数试剂盒-8（CCK-8）分析GPRC5A基因敲除小鼠气管上皮细胞对化疗药物的敏感性。采用皮下肿瘤形成实验探讨下调ABCB1表达是否可抑制体内肺腺癌增殖。采用免疫荧光和免疫沉淀实验研究GPRC5A和ABCB1之间潜在的调控关系。

结果 ABCB1在肺腺癌细胞系和人类肺腺癌组织中表达上调。GPRC5A基因敲除小鼠的气管上皮细胞及肺组织的ABCB1表达高于野生型小鼠。与GPRC5A野生型小鼠的气管上皮细胞相比，GPRC5A基因敲除小鼠的气管上皮细胞对塔立奇达和多柔比星更敏感。注射移植细胞28天后，接受ABCB1基因敲除细胞移植的GPRC5A-/- C57BL/6小鼠的肺肿瘤的体积和重量均明显低于野生型细胞移植小鼠（P = 0.0043，P = 0.0060）。此外，免疫荧光和免疫沉淀实验表明，GPRC5A通过直接结合方式调控ABCB1的表达。

结论 GPRC5A通过抑制ABCB1表达降低肺腺癌增殖。GPRC5A调节ABCB1表达的途径有待研究。

关键词: ATP 结合盒 B 亚家族成员 1, G蛋白偶联受体家族C5组成员A, 肺腺癌, 小鼠

Abstract:

Objective Aberrant expression of ATP binding cassette subfamily B member 1 (ABCB1) plays a key role in several cancers. However, influence of G protein coupled receptor family C group 5 type A (GPRC5A)-regulated ABCB1 expression on lung adenocarcinoma proliferation remains unclear. Therefore, this study investigated the effect of GPRC5A regulated ABCB1 expression on the proliferation of lung adenocarcinoma.

Methods ABCB1 expressions in lung adenocarcinoma cell lines, human lung adenocarcinoma tissues, and tracheal epithelial cells and lung tissues of GPRC5A knockout mice and wild-type mice were analyzed with RT-PCR, Western blot, or immunohistochemical analysis. Cell counting kit-8 assay was performed to analyze the sensitivity of tracheal epithelial cells from GPRC5A knockout mice to chemotherapeutic agents. Subcutaneous tumor formation assay was performed to confirm whether down-regulation of ABCB1 could inhibit the proliferation of lung adenocarcinoma in vivo. To verify the potential regulatory relationship between GPRC5A and ABCB1, immunofluorescence and immunoprecipitation assays were performed.

Results ABCB1 expression was up-regulated in lung adenocarcinoma cell lines and human lung adenocarcinoma tissues. ABCB1 expression in the tracheal epithelial cells and lung tissues of GPRC5Adeficient mice was higher than that in the wild type mice. Tracheal epithelial cells of GPRC5A knockout mice were much more sensitive to tariquidar and doxorubicin than those of GPRC5A wild type mice. Accordingly, 28 days after injection of the transplanted cells, the volume and weight of lung tumor in ABCB1knockout cell-transplanted GPRC5A^-/-C57BL/6 mice were significantly smaller than those in wild type cell-transplanted mice (P= 0.0043, P= 0.0060). Furthermore, immunofluorescence and immunoprecipitation assays showed that GPRC5A regulated ABCB1 expression by direct binding.

Conclusion GPRC5A reduces lung adenocarcinoma proliferation via inhibiting ABCB1 expression. The pathway by which GPRC5A regulates ABCB1 expression needs to be investigated.

Key words: ATP binding cassette subfamily B member 1, G protein coupled receptor family C group 5 type A, lung adenocarcinoma, mice

基金资助: 国家自然科学基金(82103571);山东省自然科学基金青年项目(ZR2021QH295)

Yun Li, Wen-Wen Cui, Zhong-Fa Yang, Wen-Hao Liu, Mao-Wang Bian, Jiong Deng, Tong Wang. Influence of GPRC5A-Regulated ABCB1 Expression on Lung Adenocarcinoma Proliferation[J].Chinese Medical Sciences Journal, 2024, 39(1): 9-18.

图/表 5

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