不同转移表型肺腺癌细胞磷脂酰肌醇的合成

Fidler于七十年代从B16癌细胞系分离出转移性高低不同的细胞亚群。又证实某些信息分子可使其互变。许多学者对高转移性和低（或不）转移性癌细胞的酶、脂类等进行了比较,但迄今为止,对两者生物学特性的差异尚无定论。本文通过同位素标记法,对高转移性及低（或不）转移性肺腺癌细胞磷脂酰肌醇（PI）的合成进行了研究。证明两者有明显差异。

Characterization of Two Human Lung Adenocarcinoma Cell Lines by Reciprocal Chromosome Painting

Lung cancer is a leading cause of cancer death worldwide. Some lung cancer patients correlate with a gas of radon besides smoking. To search for common chromosomal aberrations in lung cancer cell lines established from patients induced by different factors, a combined approach of chromosome sorting, forward and reverse chromosome painting was used to characterize karyotypes of two lung adenocarcinoma cell lines： A549 and GLC-82 with the latter line derived from a patient who has suffered long-term exposure to environmental radon gas pollution. The chromosome painting results revealed that complex chromosomal rearrangements occurred in these two lung adenocarcinoma cell lines. Thirteen and twenty-four abnormal chromosomes were identified An A549 and GLC-82 cell lines, respectively. Almost half of abnormal chromosomes in these two cell lines were formed by non-reciprocal translocations, the others were derived from deletions and duplication/or amplification in some chromosomal regions. Furthermore, two apparently common breakpoints, HSA8q24 and 12q14 were found in these two lung cancer cell lines.

Enhanced chemosensitivity of p73α gene transferred into H1299 cell line of human lung adenocarcinoma

Objective: To study the effects of transferred wild type p73α gene on the sensitivity to the chemotherapeutic agents and the growth of p53-null H1299 cells of human lung adenocarcinoma. Methods: The pcDNA3-HA-p73α plasmid was transferred into the cultured p53-null H1299 cells of human lung adenocarcinoma with the mediation of Dosper liposome; The cells resistant to G418 were selected. The expression of p73α gene in the cells was examined with Western blot. MTT assay was used to analyze the response of the transfected cells to cis-dichlorodiamine platinum (cDDP) and adriamycin (ADM). The rate of drug-induced apoptosis of the transfected cells was determined with flow cytometry and DNA fragmentation assay. The changes of the biological behaviors were observed with colony formation assay. Results: The transfected H1299 cells of human lung adenocarcinoma over-expressed p73α protein stably. MTT assay showed that the IC 50 values of cDDP and ADM were reduced by approximately 7 fold and 130 fold respectively in the transfected cells as compared with the untransfected ones. Lower concentration of the chemotherapeutic agents (1.25 μmol/L of cDDP and 0.05 μmol/L of ADM) could be employed to suppress markedly the growth of the transfected H1299 cells. The apoptotic rate induced by cDDP was increased from 10.1% to 38 4% (P<0.01) and that of ADM from 12.1% to 49.3% (P<0.01). The clonogenecity after the administration of chemotherapeutic agents was significantly lower in the transfected H1299 cells than in the parental cells (P<0.01). The sensitive enhancement ratios were 1.8 and 2.6 for cDDP and ADM respectively. Conclusion: The transfection of H1299 cells with wild type p73α gene results in an increase of the sensitivity of the cells to chemotherapeutic agents.

Synergistic effects of hyperthermia and cisplatin on human lung adenocarcinoma cell line H1299

Objective： To observe the synergistic effects of hyperthernia and cisplatin on human lung adenocarcinoma cell line H 1299. Methods： Cells treated with or without hyperthermia at 42 ℃ for 2 h were detected cisplatin sensitivity with CCK-8 assay. After managed with hyperthermia at 42 ℃ for 2 h, ciplatin 10 μg/mL for 24 h or both of two sides, inhibition ratio was detected with CCK-8 assay and flow cytometry analyzed DNA contents in every phase of cell proliferation. Results： After hyperthermia, IC50 values of cell to cisplatin decreased from 9.51 μg/mL to 6.07 μg/mL. CCK-8 assay showed hyperthermia and ciplatin had synergistic killing effects, and FCM also showed hyperthermia had significant effects on cell cycle with increase of Go/G1 phase population and reduction of S phase population. Conclusion： Hyperthermia can significantly increase the sensitivity of H1299 cell line to cisplatin.

Effects of herpes simplex virus thymidine kinase/acyclovir system on growth of human pulmonary adenocarcinoma A549 cell line in vitro and in vivo

Objective: To observe the effect of anciclovir (ACV) treatment on tumors induced by inoculation of TK gene-transfected human pulmonary adenocarcinoma A549 cells in nude mice. Methods: A recombinant plasmid containing TK gene was constructed and transfected into A549 cells by electroporation. The sensitivity of the transgenic cells (A549-TK) to ACV was examined by MTT assay in vitro and for in vivo observation, inoculation of A549-TK and A-549 cells into nude mice was separately performed to induce tumor growth, the response of which to ACV treatment was observed, and the tumor tissues were pathologically examined. Results: A recombinant plasmid containing TK gene was successfully constructed and transfected into A549 cells. The sensitivity of A549-TK cells to ACV was 43 times higher than that of A549 cells. The tumors induced by A549-TK cells showed no significant increase in size after ACV treatment (P>0. 05) , and light microscopy revealed local tissue necrosis, karyoklasis, and nuclei disappearance. Conclusion: A549-TK cells acquires sensitivity to ACV both in vitro and in vivo, and ACV can inhibit the growth of tumors induced by A549-TK cell inoculation in nude mice.

Genome-wide identification of cancer-related polyadenylated and non-polyadenylated RNAs in human breast and lung cell lines

Eukaryotic mRNAs consist of two forms of transcripts:poly(A)+ and poly(A),based on the presence or absence of poly(A) tails at the 3 end.Poly(A)+ mRNAs are mainly protein coding mRNAs,whereas the functions of poly(A) mRNA are largely unknown.Previous studies have shown that a significant proportion of gene transcripts are poly(A) or bimorphic(containing both poly(A)+ and poly(A) transcripts).We compared the expression levels of poly(A) and poly(A)+ RNA mRNAs in normal and cancer cell lines.We also investigated the potential functions of these RNA transcripts using an integrative workflow to explore poly(A)+ and poly(A) transcriptome sequences between a normal human mammary gland cell line(HMEC) and a breast cancer cell line(MCF-7),as well as between a normal human lung cell line(NHLF) and a lung cancer cell line(A549).The data showed that normal and cancer cell lines differentially express these two forms of mRNA.Gene ontology(GO) annotation analyses hinted at the functions of these two groups of transcripts and grouped the differentially expressed genes according to the form of their transcript.The data showed that cell cycle-,apoptosis-,and cell death-related functions corresponded to most of the differentially expressed genes in these two forms of transcripts,which were also associated with the cancers.Furthermore,translational elongation and translation functions were also found for the poly(A) protein-coding genes in cancer cell lines.We demonstrate that poly(A) transcripts play an important role in cancer development.