Application of Antimicrobial Peptides in Preparation of Drugs for Treating Lung Cancer
A technology for therapeutic drugs and antibacterial peptides, used in drug combinations, anti-tumor drugs, pharmaceutical formulations, etc., to achieve the effects of small molecular weight, improved therapeutic effect, hemolytic activity and low cytotoxicity
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Embodiment 1
[0024] Example 1: cec4 inhibits the accumulation of exosomes in the culture supernatant of NCI-A549 cells in a dose-dependent manner
[0025] The inventors used a series of cecropins derived from blood-sucking insects as candidate molecules, including cecropin-TY1 derived from Yaofly (Parasites&Vectors, 2015), SibaCec derived from Banna gnats (Parasites&Vectors, 2015), and cecropins derived from Aedes aegypti. AeaeCec1, AeaeCec2, AeaeCec3, AeaeCec4 and AeaeCec4 (Parasites&Vectors, 2018), these antibacterial peptide cecropins are renumbered as cec1 to cec7, and the amino acid sequences of cec1 to cec7 are shown in sequence as SEQ ID No.1-SEQ ID No.7, above The polypeptides are all synthesized by chemical method, in which all amino acids are L-type, and the carbon ends of the amino acids from cec2 to cec7 are amidated. The effect of cec1 to cec7 on the release of exosomes from non-small cell lung cancer cells NCI-A549 was detected by Western blot.
[0026] Multiple groups of ex...
Embodiment 2
[0030] Example 2: cec4 does not inhibit the proliferation of NCI-A549 cells in the concentration range of 2.5-10 μg / mL
[0031] In order to clarify whether Cec4 inhibits the production of exosomes by affecting the proliferation of NCI-A549, the effect of cec4 on the proliferation of NCI-A549 was detected. Spread NCI-A549 cells in a 96-well plate (approximately 1×10 4 cells / well), after the cells adhered to the wall, a series of double-diluted cec4 was added, and an equal volume of PBS was added to the control wells. After culturing for 48 hours, CCK-8 was used to detect the effect of different concentrations of cec4 on the proliferation of NCI-A549.
[0032] The result is as figure 2 as shown ( figure 2 In ns, no significance, *p<0.05), when the concentration of cec4 is below 40μg / mL, it does not inhibit the proliferation of NCI-A549 cells. It was shown that cec4 (2.5-10 μg / mL) inhibited the production of exosomes in NCI-A549 cells not by inhibiting cell proliferation.
Embodiment 3
[0033] Example 3: cec4 does not induce NCI-A549 cell apoptosis in the concentration range of 2.5-10 μg / mL
[0034] In order to verify whether cec4 can induce apoptosis of NCI-A549 cells, NCI-A549 cells were plated in 24-well plates (approximately 2×10 5 cells / well), add 2.5, 5, 10 μg / mL cec4 respectively, add equal volume of PBS to the control group, culture for 48 hours, use PEAnexin V cell apoptosis detection kit (BD Pharmingen) to detect NCI-A549 cells treated differently Apoptosis.
[0035] The result is as image 3 As shown, compared with the control group (PBS, image 3 a) Compared to 2.5 μg / mL ( image 3 d), 5μg / mL ( image 3 c), 10 μg / mL ( image 3 b) cec4 did not induce apoptosis of NCI-A549 cells. It was shown that cec4 (2.5-10 μg / mL) inhibited the production of exosomes in NCI-A549 cells not by inducing apoptosis.
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