The Human Lung Cancer Drug Resistance-Related Gene BC006151 Regulates Chemosensitivity in H446/CDDP Cells

Abstract

This study aimed to investigate the mechanism by which the human lung cancer drug resistance-related gene BC006151 regulates chemosensitivity by down-regulating BC006151 expression via antisense gene transfer in H446/ C DDP cells. A retroviral vector containing the antisense BC006151 sequence was constructed and transfected into H446/ C DDP cells. Transfection of the empty vector served as a negative control. The two groups of transfected cells were treated with various chemotherapeutic agents, after which morphological changes in cell ultrastructure were compared by transmission electron microscopy, cell proliferation and chemosensitivity to particular chemotherapeutic agents were compared by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide method, the effects of chemotherapy on cell cycle and apoptosis were compared by flow cytometry, and Bcl-2 was evaluated by immunohistochemistry and Western blot analysis. Results showed that apoptotic bodylike structures were identified by transmission electron microscopy in the antisense gene-transfected cells. MTT founded that these cells exhibited a significantly lower level of proliferation than the control cells (pϽ0.01), together with a markedly increased sensitivity to various chemotherapeutic agents (pϽ0.01). Flow cytometry analysis revealed that a G1 phase arrest accounted for the reduction in proliferation in the antisense gene-transfected cells; increased apoptosis was also detected (pϽ0.01). Both immunohistochemistry and western blot analysis confirmed that Bcl-2 expression was significantly down-regulated in the antisense gene-transfected cells compared to controls. In a word, down-regulation of BC006151 can significantly inhibit proliferation and increase apoptosis of H446/ C DDP cells after chemotherapy, and this gene may play an important role in the development of multidrug resistance in lung cancer.

Src homology phosphotyrosyl phosphatase 2 mediates cisplatin-related drug resistance by inhibiting apoptosis and activating the Ras/PI3K/Akt1/survivin pathway in lung cancer cells

“…In the present study, cisplatin-induced drug-resistant cells SPC-A-1/CDDP and H446/CDDP were generated from SPC-A-1 and H446 cells, respectively (30). A high expression of SHP2 was observed in these cells.…”

“…However, the development of chemoresistance is a major cause of chemotherapy failure. Thus, combination therapy directed to an appropriate target may improve therapeutic efficacy (30).…”

Expression of MSP58 in human colorectal cancer and its correlation with prognosis

“…UICC stage of tumor is the gold standard for determining prognosis in patients with colorectal cancer. But, patients with similar stages of disease may have wide discrepancy in survival [19–23]. Several new molecular prognostic factors, such as p53 and K-RAS mutations, are being evaluated in the hope that they may contribute to better assessment of survival probability [24–26].…”

Increased sensitivity of human lung adenocarcinoma cells to cisplatin associated with downregulated contactin-1

“…The bactin forward primer was 5 0 -CCTGGCACCCAGCACAAT-3 0 , and the reverse primer was 5 0 -GGGCCGGACTCGTCATAC-3 0 . The conventional RT-PCR was performed as previously described [26]. Briefly, threestep PCR was performed according to the system manual of the PCR apparatus (GeneAmp PCR system 9700, Applied Biosystems).…”