Mechanism of Scutellariae Radix-Hedyotidis Herba Against Proliferation of Lung Adenocarcinoma A549 Cells / 中国实验方剂学杂志

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-1<italic>α </italic>（HIF-1<italic>α</italic>）， 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 thecomplete medium， 40 mg·L^-1Scutellariae Radix-Hedyotidis Herba， 10 mg·L^-1 cisplatin， and 40 mg·L^-1Scutellariae Radix-Hedyotidis Herba + 10 mg·L^-1cisplatin， 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-1<italic>α</italic> 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-1<italic>α</italic>， 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 （<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^-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 （<italic>P<</italic>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 （<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 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 （<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 D₁ 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.

Regulation of mitochondrial carrier SLC25A13 on breast cancer cell cycle in vitro / 上海交通大学学报（医学版）

Objective·To investigate the role of mitochondrial solute carrier family 25 member 13 (SLC25A13) on breast cancer development.Methods·SLC25A13 mRNA and protein expressions in invasive breast cancer tissues and normal breast tissues were from The Cancer Genome Atlas (TCGA) breast cancer dataset. Survival analysis was conducted online by Kaplan-Meier software. MCF-7 cell line was used for in vitro cell assay.Knockdown of SLC25A13 and sirtuin 2 (SIRT2) were conducted by siRNA transfection. Cell viability was measured with trypan blue exclusion. Cell cycle arrest was determined by flow cytometry. The mRNA expression of SLC25A13 and P27 were detected by quantitative PCR. The protein level of SLC25A13, P27 and SIRT2 were detected by Western blotting. Protein half-life of P27 was assessed by Western blotting after ycloheximide treatment. Results·SLC25A13 was up-regulated in invasive breast cancer tissues. High expression of SLC25A13 correlated with poor overall survival and breast cancer recurrence. SLC25A13 knockdown inhibited MCF-7 cell cycle progression. P27 and SIRT2 both accumulated after SLC25A13 knockdown. P27 accumulation resulted from prolonged protein half-life. Knockdown of SIRT2 restored cell cycle arrest as well as P27 accumulation caused by SLC25A13 silencing. Conclusion·High expression of SLC25A13 may promote cell cycle progression via SIRT2 in breast cancer development.