Cancer-associated-fibroblasts regulate the chemoresistance of lung cancer cell line A549 via SDF-1 secretion
10.3760/cma.j.issn.0253-3766.2017.05.004
- VernacularTitle: 肿瘤相关成纤维细胞通过分泌基质细胞衍生因子1调节肺癌A549细胞的化疗耐药性
- Author:
Fang ZOU
1
;
Zhihua ZHANG
2
;
Yutuo ZHANG
1
;
Jianqing ZHAO
2
;
Xiulong ZHANG
2
;
Cuiling WEN
2
;
Xianyun SONG
3
;
Waimin ZHOU
4
Author Information
1. Graduate Department of Hebei North University, Zhangjiakou 075000, China
2. Department of Respiratory Diseases, the First Attached Hospital of Hebei North University, Zhangjiakou 075000, China
3. Department of NICU, Huxi Affiliated Hospital of Jining Medical College(Shanxian Central Hospital), Heze 274300, China
4. Department of Tuberculosis, the First Hospital of Changsha, Changsha 410005, China
- Publication Type:Journal Article
- Keywords:
Lung neoplasms;
Tumor microenvironment;
Cancer-associated-fibroblasts;
Stromal cell derived factor-1;
Bcl-xL;
Drug resistance
- From:
Chinese Journal of Oncology
2017;39(5):339-343
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To investigate whether cancer-associated- fibroblasts (CAF), the key component of tumor microenvironment, regulate the chemoresistant capacity of lung cancer cell line A549 through SDF-1 secretion.
Methods:Primary cell isolation techniques was used to isolate cancer-associated-fibroblasts from lung cancer patients. MTT assay was applied to determine the proliferation and chemoresistance of A549 cells. Quantative PCR was used to detect the mRNA changes of Bcl-xL. Western blotting was used to detect the protein expression of Bcl-xL. ELISA was applied to detect the SDF-1 secretion from normal fibroblasts (NF) and CAF.
Results:CAF promoted the proliferation of A549 cells, while NF had no significant effect on them. After 72 hrs incubation, the absorbance value of A549+ CAF medium group was 0.814±0.006, significantly different from the 0.753±0.006 of the A549+ NF medium group (P<0.05). The Q-PCR assay indicated that mRNA expressions of Bcl-xL in the A549 group, A549+ NF medium group and A549+ CAF medium group were 1.00±0.11, 1.10±0.09 and 3.50±0.30, respectively, showing a significant difference between the A549+ NF medium group and A549+ CAF medium group (P<0.05). The Western blot showed that protein expressions of Bcl-xL in the A549 group, A549+ NF medium group and A549+ CAF medium group were 1.00±0.08, 1.10±0.12 and 3.10±0.25, respectively, with a significant difference between the A549+ NF medium group and A549+ CAF medium group (P<0.05). The ELISA results showed that the SDF-1 concentrations in the A549+ NF medium group and A549+ CAF medium group were 3.23±0.02 and 9.53±0.10, respectively, significantly different from each other (P<0.05). The MTT assay indicated that the absorbance values of OD of A549 group, A549+ AMD3100 group, A549+ NF medium group, A549+ NF medium+ AMD3100 group, A549+ CAF medium and A549+ CA Fmedium+ AMD3100 group were 0.43±0.03, 0.25±0.02, 0.48±0.03, 0.31±0.03, 0.72±0.06 and 0.45±0.03, respectively. The data of A549+ NF medium group was significantly different from that of A549+ CAF medium group (P<0.05).
Conclusions:Cancer-associated-fibroblasts enhance the drug resistance of A549 cells through SDF-1 secretion, upregulating the expression level of Bcl-xL through interaction with CXCR4. Our study not only illustrates that tumor microenvironment is able to enhance drug resistance of tumor, but also provides experimental evidence for the cancer-associated-fibroblasts as a potential therapeutic target for the treatment of lung cancer.
