Enhanced optical breakdown in KB cells labeled with folate-targeted silver-dendrimer composite nanodevices.

Enhanced optical breakdown of KB tumor cells folate-targeted with silver-dendrimer composite nanodevices (CNDs) is described. CNDs [(Ag(0))(25)-PAMAM_E5.(NH(2))(42)(NGly)(74)(NFA)(2.7)] were fabricated by reactive encapsulation, using a biocompatible template of dendrimer-folic acid (FA) conjugates. Preferential uptake of the folate-targeted CNDs (of various treatment concentrations and surface functionality) by KB cells was visualized with confocal microscopy and transmission electron microscopy. Intracellular laser-induced optical breakdown threshold and dynamics were detected and characterized by high-frequency ultrasonic monitoring of resulting transient bubble events. When irradiated with a near-infrared, femtosecond laser, the CND-targeted KB cells acted as well-confined activators of laser energy, enhancing nonlinear energy absorption, exhibiting a significant reduction in breakdown threshold and thus selectively promoting intracellular laser-induced optical breakdown. FROM THE CLINICAL EDITOR: This study presents a novel method to selectively destroy cancer cells by combining biochemical targeting with topical laser irradiation. A human epidermoid cancer cell line was targeted with folated silver-dendrimer composite nanodevices and the labeled cancer cells were subsequently destroyed by the microbubbles generated due the enhanced energy uptake of the silver nanoparticles from the laser irradiation, as compared to unlabeled cells.

Near-IR core-substituted naphthalenediimide fluorescent chemosensors for zinc ions: ligand effects on PET and ICT channels.

Near-IR (NIR) emission can offer distinct advantages for both in vitro and in vivo biological applications. Two NIR fluorescent turn-on sensors N,N'-di-n-butyl-2-(N-{2-[bis(pyridin-2-ylmethyl)amino]ethyl})-6-(N-piperidinyl)naphthalene-1,4,5,8-tetracarboxylic acid bisimide and N,N'-di- n-butyl-2-[N,N,N'-tri(pyridin-2-ylmethyl)amino]ethyl-6-(N-piperidinyl)naphthalene-1,4,5,8-tetracarboxylic acid bisimide (PND and PNT) for Zn(2+) based on naphthalenediimide fluorophore are reported. Our strategy was to choose core-substituted naphthalenediimide (NDI) as a novel NIR fluorophore and N,N-di(pyridin-2-ylmethyl)ethane-1,2-diamine (DPEA) or N,N,N'-tri(pyridin-2-ylmethyl)ethane-1,2-diamine (TPEA) as the receptor, respectively, so as to improve the selectivity to Zn(2+). In the case of PND, the negligible shift in absorption and emission spectra is strongly suggestive that the secondary nitrogen atom (directly connected to the NDI moiety, N(1)) is little disturbed with Zn(2+). The fluorescence enhancement of PND with Zn(2+) titration is dominated with a typical photoinduced electron-transfer (PET) process. In contrast, the N(1) atom for PNT can participate in the coordination of Zn(2+) ion, diminishing the electron delocalization of the NDI moiety and resulting in intramolecular charge-transfer (ICT) disturbance. For PNT, the distinct blueshift in both absorbance and fluorescence is indicative of a combination of PET and ICT processes, which unexpectedly decreases the sensitivity to Zn(2+). Due to the differential binding mode caused by the ligand effect, PND shows excellent selectivity to Zn(2+) over other metal ions, with a larger fluorescent enhancement centered at 650 nm. Also both PND and PNT were successfully used to image intracellular Zn(2+) ions in the living KB cells.

Expression and functional characterization of the system L amino acid transporter in KB human oral epidermoid carcinoma cells.

We have examined the expression and function of system l amino acid transporter in KB human oral epidermoid carcinoma cells. The KB cells express l-type amino acid transporter 1 (LAT1) in plasma membrane, but not l-type amino acid transporter 2 (LAT2). The [14C]l-leucine uptake by KB cells is inhibited by system l selective inhibitor BCH. The majority of [14C]l-leucine uptake is, therefore, mediated by LAT1. These results suggest that the transport of neutral amino acids including several essential amino acids into the KB cells mediated by LAT1 and the specific inhibition of LAT1 in oral cancer cells will be a new rationale for anti-cancer therapy.

Intracellular calcium changes and chemosensitivities of human epidermoid carcinoma cell lines after exposure to cisplatin.

In order to elucidate the mechanisms of cisplatin (cis-diamminedichloroplatinum; CDDP)-resistant tumor cells, we previously established a CDDP-resistant KB cell line (KBrc cells) from a parental KB cell line derived from epidermoid carcinoma (KB cells). The KBrc cells were resistant to 5 kinds of platinum (Pt) drugs. Intracellular Pt concentrations in KBrc cells were lower than in KB cells. Decrease of intracellular Pt concentrations was one of the CDDP-resistant mechanisms. When we measured changes of intracellular calcium ion concentration ([Ca2+]i) during exposure to high-dose CDDP, a sustained elevation of the [Ca2+]i level was observed in the KB cells. These results suggest that the mechanisms underlying CDDP resistance involve changes in calcium channels and an alteration of calcium homeostasis in the tumor cell line.

Induction of resistance to hexadecylphosphocholine in the highly sensitive human epidermoid tumour cell line KB.

Hexadecylphosphocholine (HePC, Miltefosine) is a representative of the group of alkyl-lysophosphocholines showing remarkable antitumoral activity in in vitro experiments and in experimental animal tumour models. The epidermoid tumour cell line KB, which is highly sensitive to HePC (half-maximal growth inhibiting concentration, IC50: 1.2 microM; half lethal concentration, LC50: 2.8 microM), was slowly adapted to increasing concentrations of HePC. After 14 months, the adaptation process was stopped at a concentration of 10 micrograms/ml (23.5 microM). At this point, the KB cells tolerated high doses of HePC (IC50: 41.2 microM; LC50: 87.1 microM). The resistant cells (KBr) also showed crossresistance to the other well studied ether-lysophospholipids, Edelfosine (1-O-octadecyl-2-O-methyl-rac-glycero-3-phosphocholine, OMG-3PC; ET18OCH3) and Ilmofosine (1 S-hexadecyl-2-methoxymethyl-rac-(1-thio-3-hydroxy)propyl-3-phosphocho lin e, BM 41.440). Comparison of the KB and KBr cells showed that total lipid phosphate, ether-lipid content, vinyl-ether-lipid content, protein content as well as cholesterol content were unchanged. Furthermore, no changes were observed in the lipid composition between KB and KBr cells. Uptake of choline was also unchanged in both cells, but the uptake of D-myo-inositol was lower by a factor of two in the KBr cells. However, in KB cells, the addition of HePC induced a 50% reduction of D-myo-inositol-uptake, whereas in KBr cells inositol uptake was unchanged. Differences in HePC uptake and HePC metabolism were apparent between the KB and KBr cell lines. KBr cells showed a 3-fold lower uptake for HePC and a 3- to 4-fold faster metabolism of HePC than KB cells. However, the amount of non-metabolised HePC after 2 days of incubation with 1 microgram/ml HePC (LC50: 1.2 microgram/ml) in KB cells was 3- to 4-fold lower than the amount of HePC in KBR cells at 10 micrograms/ml (LC50: 37 micrograms/ml), indicating that KBr cells can incorporate higher amounts of HePC than KB cells without adverse effects for cell growth and viability. This seems to indicate that mechanisms other than slower uptake and faster metabolism are involved in the induction of resistance to HePC in KBr cells.

Relationship between cellular glutathione level and susceptibility to LAK killing in human pharyngeal carcinoma cell line.

We examined the relationship between cellular glutathione (GSH) level and susceptibility to lymphokine-activated killer (LAK) cell-mediated cytolysis in KB human pharyngeal carcinoma cells. Treatment of KB cells with D,L-buthionine-S,R-sulfoximine (BSO), a gamma-glutamyl cysteine synthetase blocker, resulted in decreased total intracellular GSH levels associated with increased susceptibility to LAK killing. In contrast, treatment with oxothiazolidine-4-carboxylate (OTZ, a precursor of cysteine), which is known to increase cellular GSH level, decreased the susceptibility of KB cells to LAK killing. Both agents had no effects on binding frequency of KB cells to LAK cells. These results suggest that intracellular GSH in tumor cells play a protective role against LAK mediated cytolysis, specially in the post-binding killing phase.