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Mechanism of Scutellariae Radix-Hedyotidis Herba Against Proliferation of Lung Adenocarcinoma A549 Cells / 中国实验方剂学杂志
Chinese Journal of Experimental Traditional Medical Formulae ; (24): 28-35, 2021.
Article in Chinese | WPRIM | ID: wpr-906047
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-6IL-6), phosphatidylinositol 3-kinase (PI3K),protein kinase B (Akt), p-protein kinase B (p-Akt), mechanistic target of rapamycin (mTOR), hypoxia-inducible factor-1<italic>α </italic>(HIF-1<italic>α</italic>), and Cyclin D<sub>1</sub> 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<sup>-1</sup>) Scutellariae Radix-Hedyotidis Herba groups, and low-, moderate-, and high-dose (5, 10, and 20 mg·L<sup>-1</sup>) 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<sup> </sup>complete medium, 40 mg·L<sup>-1 </sup>Scutellariae Radix-Hedyotidis Herba, 10 mg·L<sup>-1</sup> cisplatin, and 40 mg·L<sup>-1 </sup>Scutellariae Radix-Hedyotidis Herba + 10 mg·L<sup>-1 </sup>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 assayELISA) after 72 h. The mRNA expression levels of HIF-1<italic>α</italic> and Cyclin D<sub>1</sub> in each group were assayed by real-time polymerase chain reactionReal-time PCR), and the protein expression levels of PI3K, Akt, p-Akt, mTOR, HIF-1<italic>α</italic>, and Cyclin D<sub>1</sub> 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 (<italic>P</italic><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<sup>-1</sup> 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 (<italic>P<</italic>0.05). Considering the cell survival rate, the best concentration of cisplatin was set at 10 mg·L<sup>-1</sup>. Compared with the blank group, Scutellariae Radix-Hedyotidis Herba combined with cisplatin remarkably inhibited the proliferation of A549 cells in a time-dependent manner (<italic>P<</italic>0.05), and the differences between the combined medication group and the other two groups became more significant after 72 h of medication (<italic>P<</italic>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 (<italic>P<</italic>0.01). The mRNA expression levels of HIF-1<italic>α</italic> and Cyclin D<sub>1</sub> in all experimental groups were obviously lower than those in the blank group, with the most significant changes observed in the combined medication group (<italic>P</italic><0.05,<italic>P</italic><0.01). The protein expression levels of PI3K, p-Akt, mTOR, HIF-1<italic>α</italic>, and Cyclin D<sub>1</sub> in each experimental group was significantly down-regulated(<italic>P</italic><0.05,<italic>P</italic><0.01), and the levels in the combined medication group were even lower than those in the cisplatin group (<italic>P<</italic>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-1<italic>α</italic> axis.

Full text: Available Index: WPRIM (Western Pacific) Language: Chinese Journal: Chinese Journal of Experimental Traditional Medical Formulae Year: 2021 Type: Article

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Full text: Available Index: WPRIM (Western Pacific) Language: Chinese Journal: Chinese Journal of Experimental Traditional Medical Formulae Year: 2021 Type: Article