Objective: To investigate the relationship between chemosensitivity to L-OHP and expressions of multidrug resistance (MDR) associated factors in lymph node metastases (LNMs) of gastric carcinoma. Methods: The chemosensitivity to L-OHP was measured by MIT assay, and the expressions of P-gp, GST-π, P53, Survivin and Bcl-2 were determined by immunohistochemistry in 54 paired primary tumor (PT) and LNMs of gastric carcinoma. Results: The inhibition rates of LNMs cells for L-OHP were lower than those of PT (P<0.05). The expressions of P-gp, GST-π and Bcl-2 were higher in LNMs than in PT (P<0.05), and no signifi-cant difference was found in the expression of P53 and Survivin between LNMs and PT (P>0.05). Positive cor-relations among P-gp, P53 and Bcl-2 were found in PT and LNMs (r=0.3424, 0.7123, 0.4548, P<0.05). There was no significant difference in the expression of GST-π and Survivin between PT and LNMs (P>0.05). There was statistically negative correlation between inhibition rates and expression of P-gp, GST-π, and Survivin in PT (P<0.05). In LNMs, only Survivin was negatively correlated with inhibition rates of L-OHP (P<0.05). Conclu-sion: The LNMs of gastric carcinoma are heterogeneous with PT in respect to chemosensitivity to L-OHP and expression of multidrug resistance associated factors. The main factors that affect chemosensitivity to L-OHP are also significantly different between PT and LNMs. Effective adjuvant chemotherapy after surgery and re-version to multidrug resistance (MDR) of gastric carcinoma depend on targeting the metastatic lesions of gas-tric carcinoma.

IFT46 is one of the important components of intraflagellar transport complex B in Chlamydomonas reinhardtii, and plays important roles in the assembly, movement and perception of ciliary. To study its functional mechanism, a GST-tagged and an MBP-tagged prokaryotic expression plasmid, pGEX-2T-ift46 and pMAL-C2X-ift46 were constructed, respectively, by inserting ift46 into the pGEX-2T and pMAL-C2X vector, and then transformed into Escherichia coli BL21 (DE3) for protein expression. SDS-PAGE (15%) analysis results showed that the molecular weights of the fusion protein GST-IFT46 and MBP-IFT46 were 70 kDa and 86 kDa, respectively. We used the fusion protein GST-IFT46 purified by affinity adsorption purification (more than 95% purity) for immunity to New Zealand white rabbits. The 5th immune serum was collected and the antibody titer was determined to be 256 000 by ELISA. The antiserum was purified by Protein A affinity adsorption purification and immobilized MBP-IFT46 purification, and the specificity of polyclonal antibodies was evaluated by Western blotting and immunofluorescence. Results showed that the polyclonal antibody prepared could specifically and precisely bind IFT46 in C. reinhardtii, and IFT46 was mainly concentrated at basal body regions and few localized along the entire length of the flagellum as punctuated dots, which will make a foundation to further study the mechanism of IFT46 in cilia related diseases such as obesity, diabetes and polycystic kidney disease.
Algal Proteins ; biosynthesis ; immunology ; Animals ; Antibodies ; chemistry ; Blotting, Western ; Chlamydomonas reinhardtii ; chemistry ; genetics ; Electrophoresis, Polyacrylamide Gel ; Enzyme-Linked Immunosorbent Assay ; Escherichia coli ; Fluorescent Antibody Technique ; Intracellular Signaling Peptides and Proteins ; biosynthesis ; immunology ; Plasmids ; Rabbits ; Recombinant Fusion Proteins ; biosynthesis