ABSTRACT
Objective @# To investigate the effect of mitochondrial fission protein 1 (FIS1) on apoptosis and cisplatin resistance in tongue squamous cell carcinoma (TSCC) cells.@*Methods @#The squamous cell carcinoma cell lines SCC9 and CAL27 were used to detect the mRNA and protein levels of FIS1 after cisplatin treatment, the knockdown and overexpression of FIS1 of SCC9 and CAL27 with or without cisplatin treatment were accomplished through small interfering RNA (siRNA) and plasmid, respectively. The mitochondrial division state in cells was detected by mitochondrial staining, and the apoptosis state of cells was detected by TUNEL, flow cytometry and Caspase 3/7.@*Results@#FIS1 protein expression in tongue squamous carcinoma cells treated with cisplatin was increased, but the mRNA level did not change. Silencing of FIS1 expression reduced mitochondrial division and apoptosis in squamous cell carcinoma cells treated with cisplatin, whereas overexpression of FIS1 exhibited the opposite effects. The percentage of dividing mitochondria, the number of apoptotic cells and the activity of Caspase 3/7 in SCC9 and CAL27 cells were significantly different before and after modulation of FIS1 expression (P < 0.05). @*Conclusion@#FIS1 is involved in the regulation of cisplatin chemotherapy sensitivity in tongue squamous cell carcinoma and can be used as a new target for improving the sensitivity of cisplatin chemotherapy in oral squamous cell carcinoma.
ABSTRACT
Objective To observe the proliferation and apoptosis of oxaliplatin-resistant colon cancer cell lines and the expression of estrogen receptor related receptor(ERR)α when tetrasoanin was down-regulated. Methods Western blot and quantitative real-time polymerase chain reaction(qRT-PCR)were used to detect the expression of tetrasoanin and ERRα of colon cancer cells and oxaliplatin resistant cells in mRNA and protein levels. ERRα inhibitor XCT790 was used to down-regulate ERRα expression, The expression of ERRα was down regulated by ERR α inhibitor XCT790,and the level of tetrasoanin,apoptosis and proliferation of L-OHP-SW620 cells were detected by Western blot, flow cytometry and MTT.Tetrasoanin expression was down regulated by siRNA, the expression, apoptosis and proliferation of L-OHP-SW620 cells AKT, p-AKT, tetrasoanin and ERRα were detected by Western blot,qRT-PCR,flow cytometry and MTT assay.Results The expression of tetrasoanin and ERRα protein in L-OHP-SW620 cell lines were higher than those in SW620 cells (t=6.127,P<0.01,t=12.579,P<0.01),The expression of tetrasoanin mRNA in L-OHP-SW620 cell line was higher than that in SW620 cell line(t=9.085,P< 0.01). The early apoptosis rate of L-OHP-SW620 cells in XCT790 group after XCT790 inhibited ERR -αexpression was higher than that in NC group(t=3.297, P< 0.01). The survival rate of XCT790 group after 72 h culture was(45.264±6.249)%,lower than that of NC group((63.364 ± 9.472)%)(t=4.537, P<0.01). Compared with NC group,p-AKT protein was up-regulated(t=8.139,P<0.01),ERRα protein was down-regulated(t=6.452,P<0.01),the apoptosis rate was(17.541±2.317)%,lower than that in the sitetrasoanin group((32.892±3.296)%)(t=4.526,P<0.01), the survival in sitetrasoanin group after 72 h culture was(49.653 ± 5.945)%, lower than that in NC group ((67.376±7.934)%)(t=3.109,P<0.05).Conclusion Tetrasoanin down-regulation and p-AKT protein up-regulation decreases ERRα protein and OHP-resistant colon cell proliferation is decreased, apoptosis is increased and drug resistance is decreased.
ABSTRACT
Objective: To establish a small RNA library of Astragalus membranaceus, in order to understand the intake and absorption of miRNAs in human body, and to lay the foundation for pharmacological research of Astragali Radix decoction pieces (ARP). Methods: The decoction of A. membranaceus after desiccation, high temperature and microwave treatment was used as sample, and using high-throughput sequencing technology, the unknown miRNA sequence was obtained. Results: Totally 9931049 small RNA sequences were identified from ARP by the high-throughput sequencing technology. After compared with all hairpin-forming precursors in miRBase 21 database, the miRNA library of ARP decoction was established, and one conserved miRNA and 9 novel miRNA sequences were confirmed. Twelve class I small interfering RNAs (siRNAs) were obtained by further bioinformatic analysis and we found that the 5' terminal was unstable while the 3' terminal was stable thermodynamically. Conclusion: The miRNAs expression profiles of ARP is first revealed by thermodynamic treatment. Our study suggests that class I siRNA can be used as a potential biochemistry components of phytomedicine, which might contribute to the epigenetic studies on Chinese materia medica.