Construction of EGFR-MEK-TZ combined molecules and it's effects on proliferation and apoptosis of castration-resistant prostate cancer cells

doi:10.3877/cma.j.issn.1674-3253.2024.05.014

Abstract

Abstract:

Objective

To construct EGFR-MEK-TZ three combined targeting molecule LN-1 and investigate its effect on castration-resistant prostate cancer cells.

Methods

The molecular structure of LN-1 was designed and the steps of molecule synthesis were studied accordingly. Castration-resistant prostate cancer cells PC3 and LNCap were studied, then experimental group and control group were divided, respectively. The experimental group was treated with LN-1. The effects of LN-1 on the proliferative activity of PC3 and LNCap cells were detected by CCK-8 assay, clonal formation assay and EdU fluorescence staining. The expression levels of apoptosis-related genes in castration-resistant prostate cancer cells in each group were detected by RT-qPCR.

Results

The designed LN-1 molecule was composed of three active fragments: EGFR fragment, MEK fragment and alkylation fragment TZ, which were interlinked with each other through diethanolamine fragment. EGFR fragment and MEK fragment were coupled through carbonate amide bond. Using MEK fragment as starting material, the target product LN-1 with 43.3% yield was obtained. LN-1 effectively inhibited the proliferative activity of PC3 and LNCap cells (P<0.05). In addition, mRNA levels of Caspase-3, p53 and Bax in the experimental group were significantly higher than those in the control group, with significant difference (P<0.05).

Conclusions

The successful construction of EGFR-MEK-TZ three combined target molecule LN-1 suggests the rationality and feasibility of this molecular structure design, which can play an antitumor role by inhibiting proliferation and inducing apoptosis of castration-resistant prostate cancer cells PC3 and LNCap.

Key words: LN-1, Prostate cancer, LNCap, PC3, Apoptosis, EGFR, MEK, TZ

Jun Zheng, Jieying Wu, Haibo Tan, Anquan Zheng, Tengcheng Li. Construction of EGFR-MEK-TZ combined molecules and it's effects on proliferation and apoptosis of castration-resistant prostate cancer cells[J]. Chinese Journal of Endourology(Electronic Edition), 2024, 18(05): 503-508.

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References 20

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基因	正向引物	反向引物
Caspase-3	5’-GGAAGCGAATCAATGGACTCTGG-3’	5’-GCATCGACATCTGTACCAGACC-3’
Caspase-9	5’-GTTTGAGGACCTTCGACCAGCT-3’	5’-CAACGTACCAGGAGCCACTCTT-3’
Bax	5’-TCAGGATGCGTCCACCAAGAAG-3’	5’-TGTGTCCACGGCGGCAATCATC-3’
p53	5’-CCTCAGCATCTTATCCGAGTGG-3’	5’-TGGATGGTGGTACAGTCAGAGC-3’
GAPDH	5’-GTCTCCTCTGACTTCAACAGCG-3’	5’-ACCACCCTGTTGCTGTAGCCAA-3’

基因	正向引物	反向引物
Caspase-3	5’-GGAAGCGAATCAATGGACTCTGG-3’	5’-GCATCGACATCTGTACCAGACC-3’
Caspase-9	5’-GTTTGAGGACCTTCGACCAGCT-3’	5’-CAACGTACCAGGAGCCACTCTT-3’
Bax	5’-TCAGGATGCGTCCACCAAGAAG-3’	5’-TGTGTCCACGGCGGCAATCATC-3’
p53	5’-CCTCAGCATCTTATCCGAGTGG-3’	5’-TGGATGGTGGTACAGTCAGAGC-3’
GAPDH	5’-GTCTCCTCTGACTTCAACAGCG-3’	5’-ACCACCCTGTTGCTGTAGCCAA-3’

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