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To investigate the effect of microRNA-183-5p (miR-183-5p) targeting RNA-binding motif single-stranded-interacting protein 1 (RBMS1) on autophagy and migration of breast cancer cells and its mechanism.
Human breast cancer cells MCF-7 were divided into control group, NC inhibitor group (transfected with NC inhibitor) and miR-183-5p inhibitor group (transfected with miR-183-5p inhibitor), miR-183-5p inhibitor+si-NC group (transfected with miR-183-5p inhibitor and empty vector si-NC), miR-183-5p inhibitor+si-RBMS1 group (transfected with miR-183-5p inhibitor and RBMS1 knockdown plasmid si-RBMS1). The expression levels of autophagy and phosphatidylinositol-3-kinase (PI3K)/ protein kinase B (Akt)/ mammalian target of rapamycin (mTOR) pathway-related proteins in each group were detected by Western blot, and the mobility of cells in each group was detected by scratch test.
Control group, NC inhibitor group, miR-183-5p inhibitor group, miR-183-5p inhibitor+si-NC group and miR-183-5p inhibitor+si-RBMS1 group the relative expression levels of light chain 3 Ⅱ (LC3-Ⅱ)/light chain 3 Ⅰ (LC3-Ⅰ) protein were 0.29±0.03, 0.31±0.03, 0.86±0.10, 0.84±0.09 and 0.43±0.05, respectively; the relative expression levels of Beclin-1 protein were 0.18±0.02, 0.20±0.02, 0.74±0.08, 0.78±0.09 and 0.35±0.04, respectively; the relative expression levels of sequestosome-1 (P62) protein were 0.93±0.12, 0.89±0.10, 0.51±0.06, 0.54±0.06 and 0.86±0.10, respectively; the 48 h cell mobility was (62.87±7.26)%, (59.23±6.89)%, (24.13±3.49)%, (26.14±4.72)% and (40.11±5.71)%, respectively; the relative expression levels of phospho- (p-) PI3K/PI3K protein were 0.67±0.12, 0.64±0.10, 0.32±0.06, 0.35±0.06 and 0.74±0.09, respectively. The above indexes of miR-183-5p inhibitor+si-RBMS1 group were compared with those of miR-183-5p inhibitor+si-NC group, and those of miR-183-5p inhibitor group were compared with those of NC inhibitor group, and the differences were statistically significant (all P<0.05).
Inhibition of miR-183-5p expression can promote autophagy and inhibit cell migration in MCF-7 cells, while down-regulation of RBMS1 has the opposite effect, which is related to the PI3K/Akt/mTOR pathway.
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| 科 Family | 属数 Number of genus | 种数 Number of species | 占总种数比例 Percentage of total species (%) | 属 Genus | 种数 Number of species | 占总种数比例 Percentage of total species (%) |
|---|---|---|---|---|---|---|
| 鹅膏菌科Amanitaceae | 2 | 11 | 5.26 | 鹅膏菌属 Amanita | 10 | 4.78 |
| 小菇科 Mycenaceae | 2 | 12 | 5.74 | 丝盖伞属 Inocybe | 5 | 2.39 |
| 多孔菌科 Polyporaceae | 8 | 14 | 6.70 | 蜡蘑属 Laccaria | 5 | 2.39 |
| 红菇科 Russulaceae | 3 | 23 | 11.00 | 小皮伞属 Marasmius | 6 | 2.87 |
| 小菇属 Mycena | 11 | 5.26 | ||||
| 光柄菇属 Pluteus | 5 | 2.39 | ||||
| 红菇属 Russula | 17 | 8.13 | ||||
| 栓菌属 Trametes | 5 | 2.39 |
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