Yu Ya-Ya, Zhu Yan-Juan, Zou Ying, Xiao Zhen-Zhen, Shi Shuai, Liu Yi-Hong, Chang Xue-Song, Chen Ya-Dong, Zhang Hai-Bo
Department of Oncology, The Second Affiliated Hospital of Guangzhou University of Chinese Medicine, Guangzhou, China.
Department of Oncology, Guangdong Provincial Hospital of Chinese Medicine, Guangzhou, China.
Evid Based Complement Alternat Med. 2021 Oct 4;2021:9911935. doi: 10.1155/2021/9911935. eCollection 2021.
Patients with EGFR gene mutation often obtain de novo resistance to epidermal growth factor receptor tyrosine kinase inhibitors (EGFR-TKIs) or develop secondary resistance to EGFR-TKIs after taking EGFR-TKI therapy. Traditional Chinese medicine (TCM) with different treatment principles, in combination with EGFR-TKIs, plays an important role in the treatment of cancers including resistant non-small cell lung cancer (NSCLC). However, inappropriate use of TCM herbs may induce resistance to gefitinib. Therefore, it is of a great value to evaluate which TCM treatment principle should be combined with EGFR-TKIs, and which one should be avoided, and find out the potential mechanisms. The lentiviral transfection assay was used for overexpression of mutation gene in PC-9 cells to construct PC-9-PIK3CA-mutation (PC-9-PIK3CA-M) cells. Cell proliferation, apoptosis, and the expression of EGFR/PI3K/AKT and EGFR/RAS/RAF/ERK in PC-9-PIK3CA-M and H1975 cells treated by the typical drug, Qing-kai-ling (QKL) and Tan-re-qing (TRQ), or the typical drug, Shen-fu (SF) and gefitinib treatment, were detected by MTT, Annexin V/PI double labeling, and Western blot assays, respectively. Tumor xenograft and immunohistochemistry experiments were carried out to confirm the findings. PC-9-PIK3CA-M cells were less sensitive to gefitinib, when compared with PC-9 cells. QKL injection and TRQ injection, not SF injection, combined with gefitinib induced significantly increased cell growth inhibition and apoptosis in PC-9-PIK3CA-M and H1975 cells. SF injection antagonized the effect of gefitinib in promoting cancer cell apoptosis. QKL injection and TRQ injection increased the sensitivity of gefitinib by inhibiting the phosphorylation of AKT or ERK in H1975 and PC-9-PIK3CA-M cells. Similar findings were observed in H1975 xenograft mouse model. QKL and TRQ, with TCM treatment principle, should be combined with gefitinib in the treatment of NSCLC. Furthermore, drug SF should be avoided to be used together with EGFR-TKIs.
表皮生长因子受体（EGFR）基因突变的患者常常对表皮生长因子受体酪氨酸激酶抑制剂（EGFR-TKIs）产生原发性耐药，或者在接受EGFR-TKI治疗后出现继发性耐药。具有不同治疗原则的中药与EGFR-TKIs联合应用，在包括耐药非小细胞肺癌（NSCLC）在内的癌症治疗中发挥着重要作用。然而，不恰当使用中药可能会诱导对吉非替尼的耐药。因此，评估哪种中医治疗原则应与EGFR-TKIs联合应用、哪种应避免，并找出潜在机制具有重要价值。采用慢病毒转染法在PC-9细胞中过表达突变基因，构建PC-9-PIK3CA突变（PC-9-PIK3CA-M）细胞。分别通过MTT法、Annexin V/PI双染法和蛋白质免疫印迹法检测典型中药清开灵（QKL）和痰热清（TRQ）或典型中药参附（SF）与吉非替尼处理后PC-9-PIK3CA-M和H1975细胞的增殖、凋亡以及EGFR/PI3K/AKT和EGFR/RAS/RAF/ERK的表达。进行肿瘤异种移植和免疫组化实验以证实上述结果。与PC-9细胞相比，PC-9-PIK3CA-M细胞对吉非替尼不太敏感。QKL注射和TRQ注射，而非SF注射，与吉非替尼联合应用可显著增强对PC-9-PIK3CA-M和H1975细胞的生长抑制和凋亡诱导作用。SF注射拮抗了吉非替尼促进癌细胞凋亡的作用。QKL注射和TRQ注射通过抑制H1975和PC-9-PIK3CA-M细胞中AKT或ERK的磷酸化增加了吉非替尼的敏感性。在H1975异种移植小鼠模型中观察到了类似结果。具有清热化痰治疗原则的QKL和TRQ应与吉非替尼联合用于NSCLC的治疗。此外，应避免将中药SF与EGFR-TKIs联合使用。
