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Use of cabyr-a/b in promoting tumor cell sensitivity to vp16 and trail
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A tumor cell and sensitive technology, applied in the direction of antineoplastic drugs, medical preparations containing active ingredients, organic active ingredients, etc., can solve problems that have not yet been seen
Active Publication Date: 2019-03-26
BEIJING NORMAL UNIVERSITY
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[0005] So far, there is no report on the relationship between CABYR, VP16 and TRAIL
Method used
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Embodiment 1
[0056] Example 1. Silencing CABYR-a / b can promote the sensitivity of lung cancer cells to the chemotherapeutic drug VP16
[0057] This example will involve CABYR-a / b protein and caspase3 protein. Wherein, the amino acid sequence of the CABYR-a protein (i.e., a subtype CABYR protein) is shown in sequence 4 in the sequence listing; the amino acid sequence of the CABYR-b protein (i.e., b subtype CABYR protein) is shown in sequence 5 in the sequence listing; The amino acid sequence of the caspase3 protein is shown in sequence 3 in the sequence listing; cleavedcaspase3 refers to a cleavage at the 175th aspartic acid of the protein shown in sequence 3.
[0058] 1. Silencing CABYR-a / b in lung cancer cells
[0059] 1. Construct silencing vector
[0060] (1) Construction of silencing vector pGPH1 / Neo-CABYR-a / b-121
[0061] Artificially synthesize the following two single-stranded DNA molecules, CABYR-a / b-S1 and CABYR-a / b-A1, and anneal them into double strands. Then the pGPH1 / Neo v...
Embodiment 2
[0131] Example 2. Silencing CABYR-a / b can promote the sensitivity of lung cancer cells to TRAIL
[0132] In addition to CABYR-a / b protein, this embodiment also involves TRAIL protein and DR5 protein. Wherein, the amino acid sequence of TRAIL protein is shown in sequence 1 in the sequence listing; the amino acid sequence of DR5 protein is shown in sequence 2 in the sequence listing.
[0133] 1. Silencing of CABYR-a / b in NCI-H460 cells
[0134] Referring to Step 1 of Example 1, three kinds of monoclonal cells were finally obtained (into NCI-H460 cells, the silencing vectors pGPH1 / Neo-CABYR-a / b-121, pGPH1 / Neo-CABYR-a / b-303 and pGPH1 / Neo-CABYR-a / b-303 and nonsense interfering sequence).
[0135] 2. The cell survival rate of NCI-H460 cells stably silenced by CABYR-a / b was detected by MTT assay
[0136] Three kinds of monoclonal cells obtained in step 1 (into NCI-H460 cells were transferred into silencing vectors pGPH1 / Neo-CABYR-a / b-121, pGPH1 / Neo-CABYR-a / b-303 and nonsense inter...
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Abstract
The invention discloses application of CABYR-a / b to promotion of sensibility of cancer cells to VP16 and TRAIL and provides application of CABYR protein inhibitor to one of the following circumstances from (a) to (c): (a), promoting sensibility of cancer cells to VP16 and / or TRAIL; (b), preparing a product for promoting sensibility of cancer cells to VP16 and / or TRAIL; (c), preparing a product for cooperating with VP16 and / or TRAIL for treating cancers. It is proved by experiments that silent CABYR-a / b can promote DR5 protein expression in the cancer cells, breakage of caspase 3 in the cancer cells induced and caused by VP16 is enhanced, and apoptosis of cancer cells induced and caused by VP16 and / or TRAIL is enhanced. It is proved by the research result that the CABYR-a / b protein inhibitor has great significance in cooperation with VP16 and / or TRAIL for treating cancers, such as lung cancer.
Description
technical field [0001] The invention belongs to the field of biotechnology and relates to the application of CABYR-a / b in promoting the sensitivity of tumor cells to VP16 and TRAIL. Background technique [0002] Calcium-binding tyrosine phosphorylation-regulated protein (CABYR), first discovered in sperm in 2002, includes 6 transcripts encoding 5 protein subtypes, including CABYR-a,- b, -c, -d, and -e (two transcripts simultaneously encode the same isoform, CABYR-c). CABYR is testis-specific, and it is localized in the cytoplasm of spermatids and in the flagella of mature spermatozoa. According to literature reports, CABYR is expressed in various cancers, such as brain cancer, esophageal cancer, liver cancer and lung cancer. However, there are only few reports on the biological function of CABYR in liver cancer and lung cancer. For example, silencing CABYR-c in HepG2 (human liver cancer cells) can slow down the cell growth rate. The biological function of CABYR needs to b...
Claims
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