1.Expression of stomach cancer-associated transcript 16 in gastric cancer tissues and its effects on gastric cancer biological behavior
Jianfeng ZHANG ; Qianfeng ZHANG ; Xiaoling KUAI ; Zhongshang SUN ; Zhenbiao MAO
Chinese Journal of Digestion 2017;37(5):313-318
Objective To study the expression of long non-coding RNA (LncRNA) stomach cancer-associated transcript 16 (STCAT16) in gastric cancer tissues and its effects on the proliferation, migration and invasion of gastric cancer cells.Methods The different expression of STCAT16 in 32 cases of gastric carcinoma and corresponding adjacent tissues was detected by real-time fluorescence quantitative polymerase chain reaction (PCR).The STCAT16 overexpression plasmid and empty vector was separately transfected gastric cancer cell line AGS with low expression of STCAT16.The cell proliferation of empty vector group, non-transfection group and STCAT16 analogue transfection group was evaluated by cell counting kit-8 (CCK-8) assay at 0, 24, 48, 72 and 96 hours after transfection.The colony forming ability was tested by colony formation assay.The cell invasion ability was measured by Transwell chamber assay and migration ability was tested by scratch-wound assay.The effects of STCAT16 on tumorigenicity in vivo were verified by tumorigenicity experiments in nude mice.T-test and one-way analysis of variance were performed for data analysis.Repeated measures analysis of variance was used to compare the repeated measured data among groups.Chi square test and Fisher exact probability method were used for comparison of counting data.Results The expression of STCAT16 in gastric cancer tissues was low (0.87±0.19), while it was high in corresponding adjacent tissues (2.32±0.37), and the difference was statistically significant (t=-20.859, P<0.05).The expression of STCAT16 of STCAT16 analogue transfection group was higher than that of empty vector group (3.43±0.25 vs 1.00±0.06), and the difference was statistically significant (t=-16.795,P<0.05).Compared to empty vector group and non-transfection group, the cell proliferation decreased in STCAT16 analogue transfection group at 72 and 96 hours after transfection (1.41±0.07, 1.42±0.08, 1.03±0.09, and 1.72±0.11, 1.78±0.14, 1.24±0.08, respectively), and the differences were statistically significant (t=15.043,5.358, 12.193 and 8.109, all P<0.05).The results of colony formation assay indicated that the colony forming ability of gastric cells in STCAT16 analogue transfection group was lower than that in empty vector group (97.3±9.1 vs 185.0±20.1) and non-transfection group (97.3±9.1 vs 138.0±11.1), and the differences were statistically significant (t=11.634 and 4.417,both P<0.05).The results of Transwell assay showed that the number of AGS cells passing through the membrane of STCAT16 analogue transfection group was significantly less than those of empty vector group and non-transfection group (151.0±28.1 vs 228.0±38.2 and 151.0±28.1 vs 199.3±17.9), and the differences were statistically significant (t=4.823 and 4.747,both P<0.05).After transfection for 48 hours and 72 hours, the scratch-wound repair rate of STCAT16 analogue transfection group decreased, compared with those of empty vector group and non-transfection group ((52.67±6.11)%, (53.33±5.51)%, (42.67±4.72)%, and (90.67±2.51)%, (90.60±5.41)%, (69.67±1.52)%, respectively), and the differences were statistically significant (t=5.773, 5.955, 21.000 and 5.881, all P<0.05).The results of tumorigenicity in nude mice showed that compared with those of empty vector group, the tumor size of STCAT16 analogue transfection group was smaller at one-, two-, three-and four-week ((0.42±0.10) cm3 vs (0.16±0.05) cm3, (0.66±0.13) cm3 vs (0.34±0.05) cm3, (1.25±0.22) cm3 vs (0.54±0.13) cm3, (2.54±0.46) cm3 vs (0.78±0.41) cm3)), and the differences were statistically significant (t=3.175, 4.190, 7.996 and 9.705, all P<0.05).Conclusions STCAT16 is lowly expressed in gastric cancer tissues.The proliferation, migration, invasion ability and tumorigenicity in nude mice of gastric cancer cell is inhibited after upregulating the expression of STCAT16.
2.Genotype reliability of short tandem repeats typing from minute DNA.
De-jian LU ; Hong-yu SUN ; Li-xian CHEN
Journal of Forensic Medicine 2003;19(3):151-153
OBJECTIVE:
To understand the forensic practice of DNA profiling from minimal amount of DNA.
METHODS:
Serial dilutions of DNA were amplified with the PowerPlex 16 System Kit, then the genotyping of short tandem repeat(STR) was performed by ABI 377 DNA automated Sequencer.
RESULTS:
When the mount of DNA template was less than 250 pg, allelic drop-out apparently occurred at several loci. Other disturbed peaks, such as artefact bands and imbalanced heterozygote, also presented.
CONCLUSION
The anomalous results may result in incorrect genotyping. Careful and comprehensive considerations are needed to interpret the STR profile of minute DNA.
Alleles
;
DNA/genetics*
;
Forensic Medicine
;
Genotype
;
Humans
;
Male
;
Minisatellite Repeats/genetics*
;
Sequence Analysis, DNA
;
Tandem Repeat Sequences/genetics*
3.Application of single-wide-tunnel endoscopic submucosal dissection with single-clip-line traction for large early esophageal cancer and precancerous lesions
Zhongshang SUN ; Liansong YE ; Xuelian LI ; Zhiying GAO ; Zhenguo PAN ; Bing HU ; Feng PAN
Chinese Journal of Digestive Endoscopy 2024;41(10):798-804
Objective:To evaluate the clinical efficacy of single-wide-tunnel endoscopic submucosal dissection with single-clip-line traction (W-ESTD) for the treatment of early esophageal cancer and precancerous lesions with large area (≥ 3/4 circumference).Methods:A retrospective analysis was performed on patient data of large early esophageal cancer or precancerous lesions treated with digestive endoscopy at the Affiliated Huai'an NO.1 People's Hospital of Nanjing Medical University from January 2018 to January 2023. Patients were divided into W-ESTD group and endoscopic submucosal double-tunnel dissection (D-ESTD) group based on the technique used. Surgical speed, en bloc resection rate, R0 resection rate, curative resection rate, intraoperative and postoperative complications were compared between the two groups.Results:A total of 44 patients with large early esophageal cancer or precancerous lesions were included in this study, including 23 cases in the W-ESTD group and 21 cases in the D-ESTD group. There was no statistically significant difference in baseline data between the two groups ( P>0.05). The operating speeds of W-ESTD and D-ESTD groups were 29.97±11.89 mm2/min and 22.65±6.30 mm2/min, respectively, with significant difference ( t=2.580, P=0.014). There was no statistically significant difference between the two groups in terms of en bloc resection rate [95.7% (22/23) VS 100.0% (21/21), P=1.000], R0 resection rate [87.0% (20/23) VS 90.5% (19/21), P=1.000], or curative resection rate [73.9% (17/23) VS 85.7% (18/21), P=0.462]. No recurrence occurred. Intraoperative muscular injury occurred in 3 cases in the W-ESTD group and 5 cases in the D-ESTD group, and postoperative esophageal stricture occurred in 11 cases and 8 cases respectively, with no significant differences between the two groups ( P>0.05). Conclusion:Compared to D-ESTD, W-ESTD can significantly improve surgical speed and demonstrate itself as a safe and effective approach for treating large early esophageal cancer and precancerous lesions.