Establishment and characterization of a new cell line derived from human colorectal laterally spreading tumor

Abstract

CONCLUSION:We established and characterized a colorectal cancer cell line, LST-R1 and LST-R1 has an obvious malignant tendency, which maybe partially attributed to the changes of the expression of some adhesion molecules, such as E-cadherin. It is also a versatile tool for exploring the original and progressive mechanisms of laterally spreading tumor and the early colon cancer genesis. INTRODUCTIONTwo routes have been described that can lead to colon carcinogenesis: one is the adenoma-carcinoma model, which is the main carcinogenetic pathway of prudent colorectal tumor and has been widely accepted in the west. However, an increasing number of superficial or flat colorectal tumors have been reported, particular in Japan [1,2] . The superficial tumors are considered to have a distinct clinicopathologic, genetic feature and behave a different carcinogenetic pathway, called de novo pathway [3][4][5][6] . Laterally spreading tumor (LST) is a unique subtype of superficial colorectal tumor, which was first reported by Kudo S in 1993 [7,8] and many names such as granular cluster type, nodule-aggregating tumor, creeping tumor, superficial spreading (epithelial) tumor, flower-bed-like lesion had been used. In 1998, it was defined as tumors originated from the colorectal mucous membranes and mainly extends laterally rather than vertically with its diameter greater than 1 cm ( Figure 1A) [9][10][11][12] . It has a tendency to affect the rectum, sigmoid colon, and caecum. In some studies, it has been reported that the carcinogenesis rate of LST is 8.64%-52.5% [13][14][15] . Abstract AIM: To study the molecular mechanism of laterally spreading tumor (LST), a cell line [Laterally Spreading Tumor-Rectum 1 (LST-R1)] was derived and the characteristics of this cell line were investigated. BASIC RESEARCH METHODS:A new cell line (LST-R1) originated from laterally spreading tumor was established. Properties of the cell line were characterized using scanning and transmission electron microscopy, immunohistochemistry method, cytogenetic analysis and nude mice xenograft experiments. In vitro invasion assay, cDNA microarray and Western blotting were used to compare the difference between the LST-R1 and other colorectal cancer cell lines derived from prudent colon cancer. RESULTS:Our study demonstrated that both epithelial special antigen (ESA) and cytokeratin-20 (CK20) were expressed in LST-R1. The cells presented microvilli and tight junction with large nuclei. The karyotypic analysis showed hyperdiploid features with structural chromosome aberrations. The in vivo tumorigenicity was also demonstrated in nude mice xenograft experiments. The invasion assay suggested this cell line has a higher invasive ability. cDNA microarray and Western blotting show the loss of the expression of E-cadherin in LST-R1 cells.

Glutamatergic Excitation and GABA Release from a Transplantable Cell Line

“…Thus, it is likely that after a longer period in vitro, the rate of proliferation will slow down, because this cell line has not been reported to result in tumor formation (10,11). Cell line RMNE6 was also immortalized using the temperature-sensitive mutant of the SV40 large antigen, and it shows a fourfold decrease in cell proliferation (raw data) between 33°C and 39.5°C after 6 days in vitro (41), comparable to what we observed with cell line M213-2O CL-4.…”

“…In another study, Thompson found that cell line CN 1.4 releases 30% of its total GABA content after a high K + stimulus (38). Wang et al (41) reported on another GABA-producing cell line, RMNE6, established from E13 rat ventral mesencephalon cells, that has 4.13 ± 0.31 μmol/l GABA in the cytoplasm. Cell line M213-2O CL-4 has 239.68 μmol/g protein of intracellular GABA, and after high potassium stimulation it releases 1.7% (4.12 μmol/g protein) of this GABA.…”

Overexpression of hTERT in potentially malignant colorectal laterally spreading tumors

“…It has been reported that tumor cell lines Lan-1, HeLa and Co115 are telomerase-positive (27). A study describing the establishment of the LST cell line demonstrated that the histopathology of a nude mouse xenograft tumor was a poorly differentiated adenocarcinoma and cells cultured from the xenograft tumor had the same properties as the original cell line (28). Thus, the LST cell line has higher malignancy than normal adenomas.…”

Cost effectiveness of pharmacogenetic testing for uridine diphosphate glucuronosyltransferase 1A1 before irinotecan administration for metastatic colorectal cancer

“…The primary normal colon epithelium cells, human CRC cell lines LST,20 SW1116, HCT1116, HT29, SW480, SW620 and LoVo were cultured in RMPI containing 10% FBS. Total cell lysates were extracted by PBSTDS (100 ml 10 × PBS, 10 ml 100% Triton X 100, 5 g nadeoxycholate, 1 g SDS, 1 nM EDTA, up to 1,000 ml/day H 2 O) on ice and were centrifuged at 13,000 rpm at 4°C for 15 min.…”