<p>Protein tyrosine kinase 6 (<i>PTK6</i>/<i>BRK</i>) is implicated in tumor cell survival and has emerged as a potential therapeutic target in pancreatic cancer. Here, the aim of this study was to evaluate the cationic nanoliposomes loaded with asymmetric PCR product (CNLAPCRP) targeting <i>PTK6</i> in pancreatic cancerous cells (PANC-1) than on pancreatic non-cancerous cells (HPDE-C7). Here, an antisense single‑stranded DNA complementary to the <i>PTK6</i> coding sequence was generated by asymmetric PCR, purified, and encapsulated into cationic nanoliposomes. Their particle size, morphology, and surface charge were characterized. Both PANC-1 and HPDE-C7 cells were exposed to CNLAPCRP (0, 0.1, 1, and 10&#xa0;µg/mL), Asymetric PCR product alone (0, 0.1, 1, and 10&#xa0;µg/mL), nanoliposome alone, and PBS as negative control for 24, 48, and 72&#xa0;h. Finally, the cell viability (MTT), apoptosis (flow cytometry), and western bloting were assessed done for all study groups. The CNLAPCRPs were near-spherical (30–120&#xa0;nm; mean ~ 60&#xa0;nm) with a zeta potential of ~ + 25 mV. Here, we found that CNLAPCRP effectively reduced PANC-1 cell viability, induces apoptosis, and decreased ptk6 protein level. The most important finding was that CNLAPCRP had a much greater effect on cancerous cells (PANC-1) than on non-cancerous cells (HPDE-C7), indicating targeted manner of CNLAPCRP. While these data support the biological activity of the CNLAPCRP, direct demonstration of <i>PTK6</i> knockdown and inclusion of sequence‑matched and formulation controls are needed to confirm target-specific gene silencing and to delineate the mechanism. Further in‑depth characterization and in vivo evaluation are warranted.</p>

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The cationic nanoliposomes loaded with asymmetric PCR product targeting PTK6 mRNA and induce apoptosis in PANC-1 cells

  • Reyhane Fooladi Vayqan,
  • Maliheh Entezari,
  • Raziye Tajali,
  • Neda Zali,
  • Ali Jebali,
  • Amir Sadeghi,
  • Mehrdad Hashemi

摘要

Protein tyrosine kinase 6 (PTK6/BRK) is implicated in tumor cell survival and has emerged as a potential therapeutic target in pancreatic cancer. Here, the aim of this study was to evaluate the cationic nanoliposomes loaded with asymmetric PCR product (CNLAPCRP) targeting PTK6 in pancreatic cancerous cells (PANC-1) than on pancreatic non-cancerous cells (HPDE-C7). Here, an antisense single‑stranded DNA complementary to the PTK6 coding sequence was generated by asymmetric PCR, purified, and encapsulated into cationic nanoliposomes. Their particle size, morphology, and surface charge were characterized. Both PANC-1 and HPDE-C7 cells were exposed to CNLAPCRP (0, 0.1, 1, and 10 µg/mL), Asymetric PCR product alone (0, 0.1, 1, and 10 µg/mL), nanoliposome alone, and PBS as negative control for 24, 48, and 72 h. Finally, the cell viability (MTT), apoptosis (flow cytometry), and western bloting were assessed done for all study groups. The CNLAPCRPs were near-spherical (30–120 nm; mean ~ 60 nm) with a zeta potential of ~ + 25 mV. Here, we found that CNLAPCRP effectively reduced PANC-1 cell viability, induces apoptosis, and decreased ptk6 protein level. The most important finding was that CNLAPCRP had a much greater effect on cancerous cells (PANC-1) than on non-cancerous cells (HPDE-C7), indicating targeted manner of CNLAPCRP. While these data support the biological activity of the CNLAPCRP, direct demonstration of PTK6 knockdown and inclusion of sequence‑matched and formulation controls are needed to confirm target-specific gene silencing and to delineate the mechanism. Further in‑depth characterization and in vivo evaluation are warranted.