To investigate the regulatory effects of aloperin on bladder cancer cell growth and endoplasmic reticulum stress-dependent mitochondrial pathway.
253J cells were randomly divided into control group, experimental-L group (25.0 μmoL·L-1 aloperin), experimental-M group (50.0 μmoL·L-1 aloperin), experimental-H group (100.0 μmoL·L-1 aloperin), 4-PBA group [100.0 μmoL·L-1 aloperin+20 μmoL·L-1 4-phenylbutyric acid (4-PBA)], si-NC (transfected with si-NC+100.0 μmoL·L-1 aloperin), si-CHOP group [transfected with si-CCAAT-enhancer-binding protein homologous protein (CHOP)+100.0 μmoL·L-1 aloperin]. Cell proliferation was detected by 5-ethynyl-2′-deoxyuridine (EdU) assay; protein expression was detected by Western blot assay; apoptosis was detected by flow cytometry; mitochondrial membrane potential (MMP) was detected by JC-1 assay.
The EdU positive cell rates in control group, experimental-L group, experimental-M group and experimental-H group were (37.51±2.98)%, (29.52±2.19)%, (23.10±2.58)% and (12.94±2.52)%, respectively; the expression levels of C/EBP homologous protein (CHOP) were 0.29±0.04, 0.53±0.05, 0.71±0.08, 0.90±0.12, respectively. The apoptosis rates in control group, experimental-H group and 4-PBA group were (4.79±0.62)%, (23.01±2.03)% and (13.75±1.14)%, respectively. The expressions of CHOP protein in control group, experimental-H, si-NC and si-CHOP groups were 0.30±0.03, 0.92±0.08, 0.91±0.11 and 0.20±0.02, respectively; the apoptosis rates were (4.39±0.30)%, (22.11±1.82)%, (22.65±2.40)% and (15.32±1.19)%, respectively; mitochondrial cytochrome C (Cyto-C) protein levels were 0.78±0.07, 0.43±0.05, 0.42±0.05 and 0.65±0.06; cytoplasmic Cyto-C protein levels were 0.51±0.06, 0.91±0.10, 0.88±0.05 and 0.61±0.04; MMP levels were (100.00±2.79)%, (57.49±3.34)%, (54.09±4.32)% and (69.91±6.81)%, respectively. The above indexes in the experimental-L, -M, -H groups were compared with the control group, the above indexes in the 4-PBA group were compared with the experimental-H group, and the above indexes in the si-CHOP group were compared with the si-NC group, the differences were statistically significant (all P<0.01).
Aloperin can inhibit the proliferation of 253J cells by up-regulating the endoplasmic reticulum stress-dependent mitochondrial apoptosis pathway mediated by CHOP.
| 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 |