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Application of knockdown lncbcas1-4_1 cell line combined with active vitamin D in the preparation of antitumor drugs

A technology of anti-tumor drugs and vitamins, applied in the field of biomedicine, can solve problems such as high recurrence rate, poor prognosis, and rapid diffusion

Active Publication Date: 2022-04-22
SUZHOU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although great progress has been made in the research on tumor treatment methods including surgery, radiotherapy and chemotherapy, there are still many problems in the treatment of cancer patients.
For example, the overall survival time of lung cancer patients has not been significantly improved, and metastatic lesions appear in advanced lung cancer; ovarian cancer patients are difficult to diagnose in the early stage, with rapid disease progression, rapid spread, high recurrence rate, and poor prognosis.

Method used

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  • Application of knockdown lncbcas1-4_1 cell line combined with active vitamin D in the preparation of antitumor drugs
  • Application of knockdown lncbcas1-4_1 cell line combined with active vitamin D in the preparation of antitumor drugs
  • Application of knockdown lncbcas1-4_1 cell line combined with active vitamin D in the preparation of antitumor drugs

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0047] lncBCAS1-4_1 is located on chromosome 20q13.2, with a full length of 380nt (52781087to52779119, Genomeassembly: hg19), which belongs to intronic non-coding RNA, and its sequence is as follows:

[0048] GGGAGGATGATGGTCACTCCAGTCGAGCTGCACAAGAGCCTCAACACCAAGGTCTGGCAGGACCACACTCTGGCCTGGGACACCATTTTCAAATCAGTCAAAGCTTGTATCGACAACCGGTTAGAGAAGTATTCTCAGCAGCCTAGTGCAGATTTCCTTTGTGACATTTATCACCAGAATCGGCTTTCAAAGAAAGAATTGTATGCTGCTGTCACAGAGCTCCAGCTGGCTGCGGTGGAAACGGTAAGGGTGGGCTGGATTTGAGTTTGCTAAATGAACGGTCTGTACCATTTTTAGATGACAGATTTTAAATAGTAATTCATCTCATTCATTTCACTGGGATTCACTAGCATCAACTCCACAAAAATGAAAAGTCTCAT;SEQ ID NO.1。

[0049] lncBCAS1-4_1 contains two exons, Exon1 is between 52780991-52781087, Exon2 is between 52779119-52779401, and the predicted score by CPC is 2.19985, suggesting that lncBCAS1-4_1 may have coding ability, and may encode 85 amino acids, amino acid sequence as follows:

[0050] MMVTPVELHKSLNTKVWQDHTLAWDTIFKSVKACIDNRLEKYSQQPSADFLCDIYHQNRLSKKELYAAVTELQLAAVETVRVGWI; SEQ ID NO. 2.

[...

Embodiment 2

[0053] Example 2 Cell Culture

[0054] (1) Cell culture conditions

[0055] A549 and OVCAR-8 cells were cultured with RPMI-1640 medium containing 10% FBS without double antibody. Calu-1 and SKOV-3 cells were cultured with RPMI-1640 medium containing 10% FBS and 1% penicillin and streptomycin. All cells were placed at 37°C in 5% CO 2 in a sterile culture environment.

[0056] (2) Cell replacement

[0057] Preheat the culture medium and PBS to room temperature, place the petri dish in an ultra-clean bench, discard the original culture medium, add 1ml of PBS, shake the petri dish gently, and discard. After washing with PBS three times, add 5ml of fresh culture base to a 60mm-diameter petri dish, and put it into an incubator to continue culturing. Each cell type was changed individually to avoid cross-contamination between cells.

[0058] (3) Cell passage

[0059] When the cell confluency reaches 80%-90%, preheat the medium, PBS and trypsin to room temperature, discard the ...

Embodiment 3

[0066] Embodiment 3 siRNAs transfection

[0067] (1) For the silencing of lncBCAS1-4_1, three interference sequences were designed, namely si-BCAS1, si-BCAS2 and si-BCAS3.

[0068] Using siRNAs freeze-dried powder and riboFECT developed by Guangzhou Ruibo Biotechnology Co., Ltd. TM CP transfection reagent. The freeze-dried powder of siRNAs was centrifuged briefly, dissolved in DEPC water, prepared into a storage solution with a concentration of 20 μM, aliquoted, and stored at -20°C. Three interference sequences were designed for lncBCAS1-4_1, namely si-BCAS1 (GCTAAATGAACGGTCTGTA; SEQ ID NO.3), si-BCAS2 (GCTGCGGTGGAAACGGTAA; SEQ ID NO.4), and si-BCAS3 (GGATTCACTAGCATCAACT; SEQ ID NO.5). riboFECT TM CP Transfection Reagent contains riboFECT TM CP Buffer(10×) and riboFECT TM CP Reagent two reagents. Among them, riboFECT TMCP Buffer (10×) needs to be diluted to 1× with PBS before use, and it is ready-to-use.

[0069] A549 / OVCAR-8 cells with a cell confluence of 80% and in ...

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Abstract

The invention discloses the application of knocking down lncBCAS1‑4_1 cell line in combination with active vitamin D in the preparation of antitumor drugs, and belongs to the technical field of biomedicine. In order to evaluate whether lncBCAS1‑4_1 affects the antitumor effect of active vitamin D, the present invention uses 1α,25(OH) 2 D. 3 Tumor cell models dealing with lncBCAS1‑4_1 deficiency and gain of function. CCK‑8 and plate cloning experiments, scratch experiments and Transwell experiments showed that reducing lncBCAS1‑4_1 significantly promoted the antitumor effect of active vitamin D, and overexpressing lncBCAS1‑4_1 significantly resisted the antitumor effect of active vitamin D.

Description

technical field [0001] The invention relates to the technical field of biomedicine, and more specifically relates to the application of knockdown lncBCAS1-4_1 cell line combined with active vitamin D in the preparation of antitumor drugs. Background technique [0002] Tumor is one of the most dangerous diseases that threaten human life and health. Globally, tumor has become the second leading cause of death, and the number of people who die from cancer each year greatly exceeds human immunodeficiency virus / acquired immunodeficiency syndrome, tuberculosis and malaria Sum. According to statistics from the International Agency for Research on Cancer in 2018, the global cancer incidence and mortality rates have increased significantly, with 18.1 million new cases and 9.6 million deaths, respectively. Lung cancer has the highest morbidity and mortality among all cancers. Due to differences in disease regions and tumor stages, the 5-year survival rate of lung cancer patients is ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K45/06A61K31/59A61P35/00C12N15/113
CPCA61K45/06A61K31/59A61P35/00C12N15/113C12N2310/10
Inventor 李冰燕王萍张增利薛雅琪裴海龙周光明玉静
Owner SUZHOU UNIV