HOTAIR/miR-1277-5p/FBN2 signaling axis is involved in recurrent spontaneous abortion by regulating the growth, migration, and invasion of HTR-8/SVneo cells†.

OBJECTIVE: This study aimed to explore the specific pathways by which HOX transcript antisense intergenic RNA contributes to the pathogenesis of unexplained recurrent spontaneous abortion. METHODS: Real-time quantitative PCR was employed to assess the differential expression levels of HOX transcript antisense intergenic RNA in chorionic villi tissues from unexplained recurrent spontaneous abortion patients and women with voluntarily terminated pregnancies. HTR-8/SVneo served as a cellular model. Knockdown and overexpression of HOX transcript antisense intergenic RNA in the cells were achieved through siRNA transfection and pcDNA3.1 transfection, respectively. Cell viability, migration, and invasion were evaluated using cell counting kit-8, scratch, and Transwell assays, respectively. The interaction among the HOX transcript antisense intergenic RNA /miR-1277-5p/fibrillin 2 axis was predicted through bioinformatics analysis and confirmed through in vitro experiments. Furthermore, the regulatory effects of the HOX transcript antisense intergenic RNA /miR-1277-5p/fibrillin 2 signaling axis on cellular behaviors were validated in HTR-8/SVneo cells. RESULTS: We found that HOX transcript antisense intergenic RNA was downregulated in chorionic villi tissues from unexplained recurrent spontaneous abortion patients. Overexpression of HOX transcript antisense intergenic RNA significantly enhanced the viability, migration, and invasion of HTR-8/SVneo cells, while knockdown of HOX transcript antisense intergenic RNA had the opposite effects. We further confirmed the regulatory effect of the HOX transcript antisense intergenic RNA /miR-1277-5p/fibrillin 2 signaling axis in unexplained recurrent spontaneous abortion. Specifically, HOX transcript antisense intergenic RNA and fibrillin 2 were found to reduce the risk of unexplained recurrent spontaneous abortion by enhancing cell viability, migration, and invasion, whereas miR-1277-5p exerted the opposite effects. CONCLUSION: HOX transcript antisense intergenic RNA promotes unexplained recurrent spontaneous abortion development by targeting inhibition of miR-1277-5p/fibrillin 2 axis.

Lenalidomide improvement of cisplatin antitumor efficacy on triple-negative breast cancer cells in vitro.

Lenalidomide is an immunomodulatory drug and possesses anti-angiogenic and immunomodulatory activities against multiple myeloma. The present study assessed the in vitro effect of lenalidomide combined with cisplatin on MDA-MB-231, a triple-negative breast cancer (TNBC) cell line and explored the underlying molecular mechanism of this combination. Cell viability, apoptosis and the protein expression of phosphorylated (p) and total extracellular signal-regulated kinase (ERK), B-cell lymphoma-2 (Bcl-2), caspase-3, cleaved poly-adenosine diphosphate-ribose polymerase (cPARP), vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (bFGF) were measured in MDA-MB-231 cells treated with different concentrations of lenalidomide, cisplatin and their combination using different biochemical assays. Lenalidomide demonstrated no significant effect on the cell viability of MDA-MB-231 cells, even at high concentrations, whereas lenalidomide in combination with cisplatin, significantly reduced cisplatin IC50 from 7.8 to 3.0 µM in MDA-MB-231 cells. In addition, lenalidomide and cisplatin in combination significantly induced cell apoptosis by 1.6- and 1.38-fold, respectively compared with lenalidomide and cisplatin alone (P<0.05). The expression levels of VEGF, bFGF and Bcl-2 proteins were significantly reduced (P<0.01), whereas caspase-3 and cleaved PARP expression were significantly increased in MDA-MB-231 cells treated with the combination compared to those treated with single agents (P<0.01). Lenalidomide treatment alone significantly reduced the p-ERK level compared with the control (P<0.05) and cisplatin treatment alone significantly increased it (P<0.01), however treatment with them in combination significantly reduced the p-ERK level in MDA-MB-231 cells compared with cisplatin treatment alone (P<0.05). In conclusion, the present study provides the basis for using lenalidomide in combination with cisplatin in TNBC therapy.

[Efficacy of cryoablation combined with CpG oligonucleotides in the treatment of murine transplanted colon carcinoma].

OBJECTIVE: To investigate the efficacy of cryoablation combined with CpG ODN in the treatment of murine transplanted colon carcinoma. METHODS: Colon carcinoma model was established by subcutaneously inoculating CT26 cells into the right flank in BALB/c mice. The tumor-bearing mice were randomly divided into 4 groups:the group of PBS injected in peritumoral area, the group of cryoablation, the group of cryoablation combined with CpG ODN, the group of CpG ODN injected in peritumoral area. The tumor size changes were measured. Serum levels of interleukin (IL)-12 and interferon-gamma(IFN-gamma) were assayed by ELISA. The rates of CD3(+)CD4(+)T, CD3(+)CD8(+)T lymphocytes in serum were counted with flow cytometry. Mice in the cryoablation group and the combined group with tumor regression were re-challenged with CT26 cells. RESULTS: The survival time of cryoablation group and combined therapy group were (80.3 + or - 5.4) days and (83.8 + or - 5.5) days, respectively, longer than (53.7 + or - 3.7) days in PBS group and (51.5 + or - 6.8) days in CpG ODN group(all P<0.05). The suppress rates of tumor cells in cryoablation group and combined therapy group were 83.8% and 86.2% respectively. After 20 days following treatment, CD3(+)CD4(+)T/CD3(+)CD8(+)T ratio and the concentrations of IL-12 and IFN-gamma in mice serum of cryoablation group and combined therapy group were higher than those in PBS group and CpG ODN group(all P<0.05). No significant difference was found in CD3(+)CD4(+)T/CD3(+)CD8(+)T ratio between cryoablation group and combined therapy group(P>0.05). However, the concentrations of IL-12 and IFN-gamma in combined therapy group were higher than those of cryoablation group(all P<0.05). After re-challenging, tumor formation rate in the cryoablation combined with CpG ODN group was 16.7%, significantly lower than that in the cryoablation group(83.8%)(P<0.05). CONCLUSION: Cryoablation combined with CpG ODN can increase antitumor immune response in mice, and therefore can decrease the tumor formation when re-challenged with CT26 cells.

[Therapeutic and immunological effects of microwave ablation combined with CpG ODN on transplanted colon carcinoma in mice].

OBJECTIVE: To investigate the therapeutic and immunological effects of microwave ablation (MA) combined with CpG ODN in mice bearing transplanted colon carcinoma. METHODS: A mouse model bearing colon carcinoma was established by subcutaneously inoculating CT26 cells into the right flank of Balb/c mice. The tumor-bearing mice were randomized into control group with PBS injection in the peritumoral area, MA group, MA combinated with CpG ODN group, and CpG ODN group with CpG ODN injection in the peritumoral area. The tumor volume changes were observed, and serum CD3(+)CD4(+) and CD3(+)CD8(+) T lmyphocytes were analyzed by flow cytometry after the treatments. Serum levels of interleukin (IL)-2, IL-12 and IFN-gamma were detected by ELISA. The mice in the MA group and the combined treatment group showing tumor regression were rechallenged with CT26 cells. RESULTS: No significant difference was found in the number of serum CD3(+), CD3(+)CD4(+), or CD3(+)CD8(+) T lymphocytes between the 4 groups. The ratio of CD3(+)CD4(+)/CD3(+)CD8(+) T lymphocytes in the combined treatment group and MA group were 1.58-/+0.10 and 1.53-/+0.13, respectively, significantly higher than that in PBS group and CpG ODN group (P<0.05). The serum concentration of IL-2, IL-12 and IFN-gamma in the combined treatment group were 64.6-/+7.4 pg/ml, 314.1-/+26.9 pg/ml and 61.9-/+7.3 pg/ml, respectively, significantly higher than those in the other 3 groups (P<0.05). The tumor formation rate in the combined treatment group was significantly lower than that in MA group (25.0% vs 75.0%, P<0.05). CONCLUSION: CpG ODN can enhance the immunity and decrease the tumor formation rate following a rechallenge with CT26 cells in mice treated with MA.