Tanshinone IIA affects the proliferation of A549/Tax by affecting the expression of MMP7 through the PI3K-AKT-mTOR signaling pathway

Abstract

Objective: This study aims to explore whether tanshinone IIA can act on paclitaxel-resistant non-small cell lung cancer A549/Tax and analyze the possible mechanisms involved.

Methods: Using the Cell Counting Kit-8 (CCK-8), we preliminarily analyzed whether tanshinone IIA has an inhibitory effect on A549/Tax cells. We utilized public datasets, self-collected transcriptome datasets, and drug target analysis to identify potential targets. We employed real-time fluorescent quantitative polymerase chain reaction (RT-qPCR) to detect the expression of core genes before and after drug treatment to analyze potential target genes and validated them using data from The Cancer Genome Atlas (TCGA). We conducted enrichment analysis on co-expressed genes of the target genes to explore potential mechanisms. Furthermore, we employed molecular docking and western blot to verify the possible mechanisms involved.

Results: The CCK8 results indicated that tanshinone IIA has a significant inhibitory effect on A549/Tax cells. The qPCR results and the analysis of TCGA data indicated that MMP7 is the target gene. Enrichment results of MMP7 co-expressed genes suggested that the PI3K-AKT signaling pathway might play a key role. Molecular docking results indicated that tanshinone IIA has strong binding activity with PI3K, AKT, mTOR, and MMP7. Western blotting results showed that tanshinone IIA might inhibit MMP7 through the PI3K-AKT-mTOR signaling pathway.

Conclusions: Tanshinone IIA may affect the proliferation of A549/Tax by influencing the expression of MMP7 through the PI3K-AKT-mTOR signaling pathway.

Keywords: Drug resistance; Lung cancer; PI3K-AKT signaling pathway; Proliferation; Tanshinone IIA.

Fig. 1

Drug effectiveness and core target screening. a Paclitaxel is effective against A549; b A549/Tax showed significant resistance to paclitaxel; c Tanshinone IIA is effective against A549; d Tanshinone IIA is effective against A549/Tax and safe against BEAS-2B; e Differentially expressed genes between lung cancer and control; f Differentially expressed genes between A549/Tax and A549; g Genes that increase or decrease sequentially in healthy controls, A549, and A549/Tax; h The intersection gene of the stepped expression gene and the target of tanshinone IIA; i PPI networks of intersecting genes; j The underlying core genes

Fig. 2

Analysis (qPCR) and validation (TCGA) of potential core genes. a Compared with A549, the expression of CCL2 in A549/Tax was increased, and tanshinone IIA increased the expression of CCL2 in A549 and A549/Tax; b Compared with A549, the expression of CTSS in A549/Tax was increased, but tanshinone IIA did not decrease the expression of CTSS in A549/Tax; c Compared with A549, the expression of MMP2 in A549/Tax was increased, but tanshinone IIA did not decrease the expression of MMP2 in A549/Tax; d Compared with A549, the expression of MMP7 in A549/Tax is increased, and tanshinone IIA can reduce the expression of MMP7 in A549 and A549/Tax; e CCL2 expression decreased in lung cancer tissues compared with normal controls; f CTSS expression decreased in lung cancer tissues compared with normal controls; g There was no significant difference in the expression of MMP2 between the control group and lung cancer; h MMP7 expression was elevated in lung cancer compared to controls

Fig. 3

Potential mechanisms of MMP7 and the value in the diagnosis and prognosis. a Co-expressed gene of MMP7; b Biological process enrichment results of co-expressed genes; c Cellular component enrichment results of co-expressed genes; d Molecular function enrichment results of co-expressed genes; e The results of KEGG enrichment of co-expressed genes suggest that the PI3K-AKT signaling pathway may play a key role; f Diagnostic value of MMP7 in lung adenocarcinoma; g Diagnostic value of MMP7 in lung squamous cell carcinoma; h The role of MMP7 in the prognosis of lung adenocarcinoma; i The role of MMP7 in the prognosis of lung squamous cell carcinoma

Fig. 4

Results of molecular docking. a Tanshinone IIA has good docking activity with PIK3R1; b Tanshinone IIA has good docking activity with AKT1; c Tanshinone IIA has good docking activity with mTOR; d Tanshinone IIA has good docking activity with MMP7

Fig. 5

Tanshinone IIA may act on A549/Tax through the PI3K-AKT signaling pathway. a Summary results of western blotting with three repeats; b Compared with the control group(A549/Tax), Tanshinone IIA and BKM120 decreased the expression of PI3K, 740Y-P could block the effect of tanshinone IIA, but Marimastat had no significant effect on the expression of PI3K; c Compared with the control group(A549/Tax), Tanshinone IIA and BKM120 decreased the expression of p-AKT, 740Y-P could block the effect of tanshinone IIA, but Marimastat had no significant effect on the expression of p-AKT; d Compared with the control group(A549/Tax), Tanshinone IIA and BKM120 decreased the expression of AKT, 740Y-P could block the effect of tanshinone IIA, but Marimastat had no significant effect on the expression of AKT; e Compared with the control group(A549/Tax), Tanshinone IIA, Marimastat and BKM120 decreased the expression of mTOR, 740Y-P could block the effect of tanshinone IIA; f The expression of MMP7 increased gradually in BEAS-2B, A549 and A549/Tax, Compared with the control group(A549/Tax), Tanshinone IIA, Marimastat and BKM120 decreased the expression of MMP7, 740Y-P could block the effect of tanshinone IIA