Raddeanin A inhibits proliferation, invasion, migration and promotes apoptosis of cervical cancer cells via regulating miR-224-3p/Slit2/Robo1 signaling pathway

Abstract

Raddeanin A (RA), an active triterpenoid saponin extracted from the Anemone raddeana regel, plays an essential role in the suppression of many malignancies. We aimed to investigate the effects and potential mechanisms of RA on cervical cancer (CC). RA was used to treat CC cell lines (Hela and c-33A) for 24 h and 48 h. Then, the invasion, migration and cell cycle distribution of these two cell lines with RA treatment were respectively detected by transwell, wound healing and flow cytometry. Results revealed that RA significantly inhibited the invasion, migration, promoted the cell cycle arrest and apoptosis of Hela and c-33A cells. Moreover, RA was confirmed to activate the Slit2/Robo1 signaling, and bioinformatics analysis and luciferase reporter assay verified that miR-224-3p could target Slit2. Additionally, miR-224-3p overexpression reversed the inhibitory effect of RA on invasion and migration of CC cells, and it also restored the promoting effects of RA on cell cycle arrest and apoptosis. Lastly, miR-224-3p-upregulation inactivated the expression of Slit2 and Robo1 in RA-treated Hela and c-33A cells. These findings demonstrated that RA inhibits proliferation, invasion, migration and promotes apoptosis of CC cells through miR-224-3p/Slit2/Robo1 signaling pathway, which might guide the future studies or treatment of this disease.

Raddeanin A Enhances Mitochondrial DNA‐cGAS/STING Axis‐Mediated Antitumor Immunity by Targeting Transactive Responsive DNA‐Binding Protein 43

“…Moreover, RA treatment dramatically upregulated cell surface expression of CRT, another known ICD marker that serve as “eat‐me” signal for tumor immune recognition (Figure 1F), further confirming the ICD‐inducing effect of RA. In line with previous report, [ 20 ] RA induced markedly apoptosis in MC38 and B16 cells (Figure S1A, Supporting Information), while blockade of apoptosis by a pan‐caspase inhibitor (Z‐VAD‐FMK), but not necroptosis inhibitor (necrostatin1) or lysosomal cell death inhibitor CA‐074 Me, inhibited RA‐induced HMGB1, ATP release and CRT surface expressions in tumor cells (Figure S1B–G, Supporting Information), indicating RA triggered ICD effects of tumor cells mainly through induction of apoptosis. To further validate the immunogenicity‐inducing effect of RA in vivo, we inoculated C57BL/6 mice with RA‐pretreated MC38 dead cells or freeze‐thawed cells on the right flank (in situ).…”

Immune-Related Genes: Potential Prognostic Factors and Regulatory Targets for Cervical Carcinoma

“…Slit2 plays a comprehensive role in the progression of tumors [43,44]. In cervical carcinoma, the expression of slit2/robo1 is abnormally low, and it plays an anticancer role [45][46][47]. These reports were consistent with our results.…”

Tetramethylpyrazine inhibits the proliferation, invasiveness and migration of cervical cancer C33A cells by retarding the Hedgehog signaling pathway

“…Wound-healing assay. C33A cells (5x10 5 cells/well) were seeded into a 6-well plate and cultured in DMEM containing 10% FBS at 37˚C until 80% confluent, and subsequently serum-starved overnight (28). A 200-µl pipette tip was used to create a wound in the cell monolayer, and the cells were then cultured in serum-free DMEM with TMP (25, 50 and 100 µM) or TMP (100 µM) + SAG.…”