Keywords

Scorpion Venom Peptide K562/A02 Babl/C Multidrug Resistance In Nude Mice

Abstract

Objective To explore the effect of peptide extract from scorpion venom (PESV) on multidrug resistance (MDR) of leukemic stem cells (LSC) in vivo. Methods K562/A02 cells were cultured, and K562/A02 stem cells were collected in the logarithmic growth phase and sorted by immunomagnetic bead method for later use; 5 of 40 BABL/c nude mice were injected with K562/A02 stem cells to form subcutaneous tumor masses for later use; A random number table method was used to divide 35 nude mice into normal control group, model group, Adriamycin (ADM)-resistant group, PESV group and ADM+PESV high, medium and low dose groups, with 5 mice in each group. The normal control group was not treated, and the remaining groups were embedded with tumor tissue. After modeling, the model group was intraperitoneally injected with 1 mL of normal saline, once a day; the ADM group was intraperitoneally injected with ADM 0.05 mg, once every other day; the PESV group was intraperitoneally injected with PESV. 2 ¦Ìg, once a day; ADM + PESV high, medium and low dose groups received intraperitoneal injection of ADM 0.05 mg, once every other day, and PESV (5, 2, 1 ¦Ìg) intraperitoneally, once a day. Administer for 14 days. Flow cytometry detects P-glycoprotein (P-gp); RT-PCR detects breast cancer resistance protein (BCRP) and multidrug resistance gene 1 (MDR1) mRNA expression; immunohistochemical method to detect aldehyde dehydrogenase 1 (ALDH1); Western blot method to detect phosphoinositide 3-kinase (PI3K) protein expression; ELISA to detect nuclear transcription factor NF- ¦ÊB content. Results The CD34+CD38- cell ratios of K562/A02 cells before and after immunomagnetic bead sorting were 31.5% and 92.8% respectively, and the drug resistance rates (IC50) were (60.33¡À10.68) ¦Ìg/mL and (58.33¡À9.72) ¦Ìg respectively. /mL, the tumor formation rate in nude mice was 100%. Compared with the model group, the differences in various detection indicators in the ADM group were not statistically significant (P>0.05); in the PESV group, the differences in BCRP, MDR1 mRNA and NF-¦ÊB factor levels were statistically significant (P<0.05); The P-gp value of the ADM+PESV high-, medium-, and low-dose groups decreased, and the PI3K protein expression was down-regulated (P<0.05). The BCRP mRNA expression of the ADM+PESV high-, medium-dose groups decreased, and the MDR1 mRNA expression increased. The differences were statistically significant. Significance (P<0.05); PI3K protein expression in the ADM+PESV high-dose group was down-regulated, and the difference was statistically significant (P<0.05). Compared with the ADM group and PESV group respectively, the levels of P-gp, BCRP mRNA, MDR1 mRNA, PI3K and NF-¦ÊB in the ADM+PESV high-dose group were significantly reduced, and the difference was statistically significant (P<0.05). There was no statistically significant difference in the ALDH1 positive rate among each group (P>0.05). Conclusion PESV combined with ADM can down-regulate the expression levels of P-gp, BCRP, MDR1, PI3K, and NF-¦ÊB in leukemia K562/A02 stem cells, enhance the sensitivity of leukemia K562/A02 stem cells to ADM in vivo, and reverse their multidrug resistance. effect

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Original research done by Liu Baoshan, Shi Zhexin, Yang Xiangdong, Yan Lili

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