Mechanism of Scutellariae Radix-Hedyotidis Herba Against Proliferation of Lung Adenocarcinoma A549 Cells
10.13422/j.cnki.syfjx.20211922
- VernacularTitle:黄芪-白花蛇舌草抑制肺癌A549细胞增殖机制
- Author:
Hai-peng SUN
1
;
Shan MA
2
;
He ZHUANG
2
;
Yue ZHAO
3
;
Ping-yi SUN
2
;
Si-yuan LIU
2
Author Information
1. Affiliated Hospital of Shandong University of Traditional Chinese Medicine（TCM），Jinan 250014，China
2. Shandong University of TCM，Jinan 250355，China
3. Second Clinical College，Shandong University of TCM，Jinan 250001，China
- Publication Type:Research Article
- Keywords:
Scutellariae Radix-Hedyotidis Herba;
lung adenocarcinoma A549 cells;
interleukin-6（IL-6）;
hypoxia-inducible factor-1α （HIF-1α）;
proliferation
- From:
Chinese Journal of Experimental Traditional Medical Formulae
2021;27(19):28-35
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To observe the effects of Scutellariae Radix-Hedyotidis Herba on the proliferation of human lung adenocarcinoma A549 cells and the expression of interleukin-6（IL-6）， phosphatidylinositol 3-kinase （PI3K），protein kinase B （Akt）， p-protein kinase B （p-Akt）， mechanistic target of rapamycin （mTOR）， hypoxia-inducible factor-1α （HIF-1α）， and Cyclin D1 at the cellular level， and to explore their molecular mechanism. Method:Following the set-up of the blank group （complete medium）， low-， moderate-，and high-dose （20， 40， and 60 mg·L-1） Scutellariae Radix-Hedyotidis Herba groups， and low-， moderate-， and high-dose （5， 10， and 20 mg·L-1） cisplatin groups， the cell were treated with the corresponding drugs for 24， 48， and 72 h for detecting their viability by tetrazolium bromide （MTT） colorimetry. A549 cells were then divided into the blank group， Scutellariae Radix-Hedyotidis Herba group， cisplatin group， and combined medication group and intervened with the complete medium， 40 mg·L-1 Scutellariae Radix-Hedyotidis Herba， 10 mg·L-1 cisplatin， and 40 mg·L-1 Scutellariae Radix-Hedyotidis Herba + 10 mg·L-1 cisplatin， respectively， for 24， 48 and 72 h， followed by the measurement of inhibitory effects against the proliferation of A549 cells in each experimental group. The level of IL-6 in cell culture supernatant was determined by enzyme-linked immunosorbent assay （ELISA） after 72 h. The mRNA expression levels of HIF-1α and Cyclin D1 in each group were assayed by real-time polymerase chain reaction （Real-time PCR）， and the protein expression levels of PI3K， Akt， p-Akt， mTOR， HIF-1α， and Cyclin D1 by Western blot. Result:After 24 h intervention， Scutellariae Radix-Hedyotidis Herba did not significantly inhibit the proliferation of A549 cells. However， 48 h later， the inhibitory effect in Scutellariae Radix-Hedyotidis Herba groups were significantly enhanced in comparison with that in the blank group （P<0.05）， exhibiting a time-dependent response. After 72 h of action， no significant change was present in the inhibitory effect of each Scutellariae Radix-Hedyotidis Herba group， so the optimal concentration of Scutellariae Radix-Hedyotidis Herba was set at 40 mg·L-1 for follow-up experiments. As demonstrated by the comparison with the blank group， cisplatin at each concentration inhibited the cell proliferation in a time-dependent manner （P<0.05）. Considering the cell survival rate， the best concentration of cisplatin was set at 10 mg·L-1. Compared with the blank group， Scutellariae Radix-Hedyotidis Herba combined with cisplatin remarkably inhibited the proliferation of A549 cells in a time-dependent manner （P<0.05）， and the differences between the combined medication group and the other two groups became more significant after 72 h of medication （P<0.01）. The IL-6 level in each experimental group， especially in the combined medication group， significantly declined in contrast to that in the blank group （P<0.01）. The mRNA expression levels of HIF-1α and Cyclin D1 in all experimental groups were obviously lower than those in the blank group， with the most significant changes observed in the combined medication group （P<0.05，P<0.01）. The protein expression levels of PI3K， p-Akt， mTOR， HIF-1α， and Cyclin D1 in each experimental group was significantly down-regulated（P<0.05，P<0.01）， and the levels in the combined medication group were even lower than those in the cisplatin group （P<0.01）. Conclusion:Scutellariae Radix-Hedyotidis Herba has an inhibitory effect on the proliferation of A549 cells， which may be related to its inhibition against the expression and secretion of IL-6/PI3K/Akt/mTOR-HIF-1α axis.
