The function and mechanism of LncRNA TTN-AS1 regulating the sensitivity of multidrug resistant osteosarcoma cells to adriamycin
Objective To investigate the function and mechanism of long non-coding RNA(LncRNA)TTN-AS1 in regulating the sensitivity of osteosarcoma cells to adriamycin(ADR).Methods Osteosarcoma cell lines were purchased from the Shanghai Cell Bank,Chinese Academy of Sciences.Reverse transcription-quantitative polymerase chain reaction(RT-qPCR)was used to detect the expression of TTN-AS1 in osteosarcoma cell line MG63 and multidrug-resistant cell line MG63/Dox.MG63/Dox was divided into Control group,si-TTN-AS1 group,ADR group,si-TTN-AS1+ADR group,si-TTN-AS1+miR-134-5p inhibitor+ADR group and si-TTN-AS1+miR-134-5p inhibitor+si-MBTD1+ADR group.The dual luciferase reporter gene experiment was used to detect the targeting relationship between TTN-AS1 and miR-134-5p,as well as the targeting relationship between miR-134-5p and malignant brain tumour domain containing 1(MBTD1).CCK8 and flow cytometry were used to detect cell proliferation and apoptosis in each group.Results The relative expression level of TTN-AS1 in MG63/Dox cells was significantly higher than that in MG63 cells(t=6.43,P＜0.05).The proliferation ability of MG63/Dox cells in 24,48,72 h in the si-TTN-AS1+ADR group was significantly lower than that in the si-TTN-AS1 group(t=4.16,5.06,5.72),and cell apoptosis was significantly higher than that in the si-TTN-AS1 group and ADR group(t=6.23,14.84),the differences were statistically significant(all P＜0.05).MiR-134-5p is the TTN-AS1 target gene,and MBTD1 is the miR-134-5p target gene.The proliferation ability of MG63/Dox cells in 24,48,72 h in the si-TTN-AS1+miR-134-5p inhibitor+ADR group was significantly higher than that in the si-TTN-AS1+ADR group(t=10.66,10.96,14.03),and cell apoptosis was significantly lower than that in the si-TTN-AS1+ADR group(t=5.58),the differences were statistically significant(all P＜0.05).The proliferation ability of MG63/Dox cells in 24,48,72 h in the si-TTN-AS1+miR-134-5p inhibitor+si-MBTD1+ADR group was significantly lower than that in the si-TTN-AS1+miR-134-5p inhibitor+ADR group(t=5.78,7.85,8.38),and cell apoptosis was significantly higher than that in the si-TTN-AS1+miR-134-5p inhibitor+ADR group(t=7.96),the differences were statistically significant(all P＜0.05).Conclusion TTN-AS1 could regulate the miR-134-5p/MBTD1 axis to affect the sensitivity of MG63/Dox to ADR.
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