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Outline
To study the regulatory effect of tripterygium glycosides（TG）on ferroptosis of osteosarcoma cells via transcription factor EB（TFEB）-lysosome pathway.
Osteosarcoma U2OS cells were cultured and divided into control group, TG groups with different concentrations（10, 20, 40, 80, 160, 320 μmol/L）, negative control（NC）group（transfected with NC siRNA）, and NC + TG group（transfected with NC siRNA combined with 160 μmol/L TG）, si-TFEB + TG（transfected with TFEB siRNA combined with 160 μmol/L TG）. Cell proliferation activity was detected by CCK-8 assay, lysosome probe Lyso-Tracker activity was detected by flow cytometry, Fe2+ and reactive oxygen species（ROS）levels were detected by enzymolysis assay. Western blot analysis was performed to detect the expression levels of TFEB in total protein and nuclear protein and the expression levels of lysosome related proteins（LAMP1, LAMP2, CTSB, CTSD）and ferroptosis related proteins（GPX4, SLC7A11）in total protein.
The survival rate of TG groups with different concentrations was lower than that of the control group（P<0.05）, and the higher the concentration of TG, the lower the survival rate. The LysoTracker activity and the expression levels of LAMP1, LAMP2, GPX4 and SLC7A11 in total protein of low, medium and high concentration TG groups（40, 80, 160 μmol/L） were lower than those in the control group, the expression levels of Fe2+, ROS and TFEB in nuclear protein, the expressions of CTSB and CTSD in total protein were higher than those in the control group（P<0.05）. The survival rate, LysoTracker activity and the expression levels of LAMP1, LAMP2, GPX4 and SLC7A11 in total protein of si-TFEB + TG group were higher than those in the NC + TG group, the expression levels of Fe2+, ROS and TFEB in nuclear protein, the expression of CTSB and CTSD in total protein were lower than those in the NC + TG group（P<0.05）.
TG affects lysosomal function by promating TFEB nuclear translocation, and then promotes ferroptosis in osteosarcoma cells.
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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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