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Outline
To investigate the effect of sodium arsenite(NaAsO2) exposure on the expression of trans-spliced tissue inhibitor of metalloproteinase 2(ts-TIMP2) in A549 cells and to analyze the role of ts-TIMP2 in arsenic-induced cell viability.
Specific primers were designed to amplify ts-TIMP2, and their specificity was validated via qRT-PCR, agarose gel electrophoresis, and DNA sequencing. CCK-8 assays were used to determine appropriate NaAsO2 concentrations and exposure duration. mRNA levels of TIMP2 and ts-TIMP2 were quantified by qRT-PCR. siRNA-mediated knockdown and lentiviral overexpression systems(carrying GFP and 3×Flag tags) were established to modulate ts-TIMP2 expression, and CCK-8 assays were performed to assess cell viability under various conditions.
The results from all three methods confirmed the specific amplification of ts-TIMP2 by these primers. NaAsO2 reduced A549 cell viability in a dose-dependent manner, with an IC50 of 48.48 μM. TIMP2 mRNA was significantly upregulated at 30 and 45 μM NaAsO2(P<0.01), while ts-TIMP2 expression increased significantly only at 45 μM(P<0.05), indicating concentration-dependent regulation. Knockdown of ts-TIMP2 decreased cell viability by 13.18% compared to the negative control(P<0.01), co-treatment with NaAsO2 further significantly suppressed cell viability(P<0.01). Conversely, ts-TIMP2 overexpression enhanced cell viability by 24.66%(P<0.01) and maintained significantly higher viability than the NaAsO2-only group even under co-treatment conditions(P<0.05).
Sodium arsenite(NaAsO2) can induce the expression of trans-spliced TIMP2(ts-TIMP2), thereby enhancing the viability of A549 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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