| Summary: | 碩士 === 中原大學 === 奈米科技碩士學位學程 === 105 === Metastasis is one hallmark of cancers responsible for 90% of cancer-related deaths. In oral cancer, ~50% of metastases have found at the prognosis declining ~30% of the 5-year survival rate. In this study, orthotopic metastatic oral cancer model was established by inoculation 6.3 x 105 SAS-G418 cells onto tongue of nude mice. The metastasis was observed to cervical area including superficial cervical lymph node (SCL), deep cervical lymph node (DCL) and submandibular gland (SG) on day 7 post SAS-G418 cells inoculation. The established metastasis model allowed to further therapeutic evaluation of lipid-platinum-chloride nanoparticles (LPC NPs). The measured LPC NPs size was 23.8±1.2nm and enhanced with outer leaflet of aminoethyl anisamide, a ligand of overexpressed sigma receptor on the surface of SAS cells for targeted therapy. The %yield of CDDP precursor was 91.6±6.7wt%, but the %encapsulation efficiency and %drug loading of LPC NPs were 2.4±0.1wt% and 89.14±8.91wt%, respectively. In vitro cell viability showed that LPC NPs significantly induced cell death (p<0.05) and promoted 2.4 or 2.8 folds stronger to reach IC50 on SAS (7±1.7µM LPC, 17±1.7µM CDDP) and SCC4 (5±4.6µM LPC, 14±1.7µM CDDP) cells, respectively.
LPC_AEAA(-) and LPC_AEAA(+) showed a significant difference (P<0.001) in animal survival rate analyzed using Kaplan-Meier survival analysis compared to CDDP or PBS group. However, LPC_AEAA(-) group showed no significant difference (P>0.05) in animal survival rate compared to LPC_AEAA(+) group and so did PBS group compared to CDDP group. In addition, the average of animal survival in LPC_AEAA(-) and LPC_AEAA(+) were significantly prolonged (P<0.001) compared to PBS (19 days), CDDP (22 days), where animal survival days for LPC_AEAA(-) and LPC_AEAA(-) were 80 and 89 days, respectively. LPC-treated groups showed significantly different (P<0.001) on IHC analysis against cell proliferation (Ki-67), tumor microvessel (CD31), DNA damage (TP53), and apoptosis (cleaved caspase-3 and TUNEL assay) markers. The protein levels of total P53 and phosphorylation of P53 on serine 15 enhanced on LPC-treated groups but varied on individual mouse. In addition, a significant induction of cleaved caspas-3 protein level was observed on LPC_AEAA(-) (P<0.05) and LPC_AEAA(+) (P<0.001) compared to CDDP or PBS group.
LPC NPs-treated groups showed no difference (P>0.05) in all parameters of liver, kidney, and muscle functions compared to PBS group, but CDDP group showed significant difference (P<0.05) on some markers including liver (aspartate aminotransferase, AST), kidney (blood urea nitrogen, BUN) and muscle (lactate dehydrogenase, LDH) functions. Results suggest that LPC NPs served minimum side effects compared to CDDP. No difference (P>0.05) activation of cytokines inflammatory factors (IL-6, IL-12 and INF-γ) were detected by enzyme-linked immunosorbent assay (ELISA). Overall, we demonstrated that LPC NPs highly improved the anticancer effect of cisplatin, prolonged animal survival rate, and could be a potential antimetastatic drug for treatment against advanced HOSCC
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