ABSTRACT
Objective To analyze the effect of trametes robiniophila on the apoptosis and the expressions of invasion and metastasis related matrix metalloproteinase 2 (MMP-2) and MMP-9 in human gastric carcinoma poorly differentiated cell line MKN-45 and medium differentiated cell line SGC-7901.Methods Human gastric carcinoma cell lines MKN-45 and SGC-7901 incubated with trametes robiniophila at the different concentrations [0 (the negative control group), 5, 10, and 20 mg/ml] for 24 h. When bifluorescently stained with acridine orange-ethidium bromide (AO-EB), morphology was observed by using microscopy. Flow cytometry was used to test the apoptosis ratio after 24, 48, and 72 h. Reverse transcription polymerase chain reaction (RT-PCR) was applied to analyze the expressions of mRNA of MMP-2 and MMP-9 after 24 h, and the absorbance values of the bands after gel electrophoresis were used as their expression levels. Results Under the fluorescence microscope, the cells of the negative control group were green (normal cells), and the membrane was even in size and integrity. The proportion of apoptotic and necrotic cells was increased with the concentration enrichment of trametes robiniophila. The apoptotic cells were yellow, and their cell membrane lost integrity. Apoptotic bodies were found in cancer cells, and cell membrane showed bud projection. The nuclei of the apoptotic cells showed brightly condensed chromatin or fragmented. Chromatin was strongly stained and located in karyotheca. The necrotic cells were dyed red with integrity of size. The apoptotic ratio of MKN-45 cell line induced by trametes robiniophila after 24 h was (6.5 ±0.8)％, (14.6±1.0)％, (18.0±1.1)％, and (23.1±1.2)％, respectively (F= 333.972, P< 0.01) in negative control group and 5, 10, and 20 mg/ml trametes robiniophila group. After 48 h, the apoptotic ratio was (7.3 ±1.2)％, (18.3 ± 1.6)％, (24.5±1.3)％, and (27.2±1.7)％, respectively (F= 528.432, P= 0.001); after 72 h, the apoptotic ratio was (7.5 ±0.9)％, (50.2 ±1.6)％, (58.0 ±1.9)％, and (69.0 ±1.4)％, respectively (F= 3814.238, P< 0.01). After SGC-7901 cell line induced by trametes robiniophila for 24 h, the apoptotic ratio was (12.9 ±1.0)％, (19.4 ± 1.2)％, (22.0±1.7)％, and (23.0±1.9)％, respectively (F= 120.190, P< 0.01) in negative control group and 5, 10, and 20 mg/ml trametes robiniophila group. For 48 h, the apoptotic ratio was (10.2 ±1.3)％, (40.9 ±1.4)％, (51.6 ±1.9)％, and (66.2 ±1.9)％, respectively (F= 1281.342, P< 0.01). For 72 h, the apoptotic ratio was (27.4 ±1.8)％, (49.7 ±1.4)％, (65.1 ±1.4)％, and (69.0 ±2.0)％, respectively (F= 1112.767, P< 0.01). The induction of apoptosis showed time and dose dependence (both P< 0.01). There was a trendency that the apoptotic ratio of SGC-7901 cell line was higher than that of the MKN-45 cell line at the same condition. RT-PCR showed that mRNA relative expression level of MMP-2 was 0.64±0.02, 0.49±0.01, 0.36±0.02, and 0.32±0.01, respectively (F= 274.321, P< 0.01) of negative control group and 5, 10, and 20 mg/ml trametes extract group after the effect of trametes extract on gastric cancer MKN-45 cell line. The relative expression level of MMP-9 in MKN-45 cell line was 0.71±0.01, 0.54±0.02, 0.47±0.02, and 0.39±0.02, respectively (F=203.948, P< 0.01). The expression level of MMP-2 in SGC-7901 cell line was 0.64±0.01, 0.42±0.02, 0.34± 0.20, and 0.29±0.01, respectively (F= 305.877, P< 0.01), while the mRNA expression level of MMP-9 was 0.65 ±0.15, 0.47 ±0.01, 0.44 ±0.01, and 0.39 ±0.02, respectively (F= 265.259, P< 0.01). Compared with the negative control group, the expression levels of MMP-2 and MMP-9 of the two cell lines were both decreased, after incubated with trametes robiniophila at the different concentrations (all P< 0.05), and with the addition of concentration, the expression levels of MMP-2 and MMP-9 were decreased. Conclusion Trametes robiniophila can induce the apoptosis of human gastric carcinoma cell lines MKN-45 and SGC-7901 in vitro and can inhibit the expressions of MMP-2 and MMP-9 and the effect of trametes robiniophila may be related with the differentiation degree.
ABSTRACT
Objective To invstigate the effect of high dose X-ray irradiation on the expression of multidrug resistance-1 (MDR1), Bcl-2, MMP7 genes. Methods A nasopharyngeal carcinoma cell line,CNE1, were irradiated with a total dose of 50 Gy. The resistance to the cisplatin of CNE1 cells and the irradiated CNE1 (CNE1 R) cells was detected by MTT. mRNAs expression of MDR1 , Bcl-2 and MMP7 was measured by quantitative RT-PCR. Results The expression of MDR1 increased in CNE1 R cells. The semiquantitative A value of MDR1 mRNA was 0.47 ±0.04, and the value of CNE1R cells (1st, 7th, 21st,28th, 35th, 42nd and 49th days after irradiation) were 0.67 ± 0. 06 (t = -5.44, P = 0. 003) ,0.70 ± 0. 01(t=-5.90,P=0. 002),0.73±0. 01(t= -6. 45,P=0. 001) ,0. 67 ± 0. 03 (t= -3.97,P=0.011),0.65 ±0.01(t = -4.43,P=0. 007),0. 62±0. 05(t= -2. 64,P=0.046) and 0.62 ±0.02(t = -3.34,P=0.021), respectively. Bcl-2 mRNA expression were 0.55 ±0.02 and 1.05 ±0.04(t = -9.93,P=0. 000) and MMP7 mRNA expression were 0.51 ±0.01 and 0.82 ±0.02(t = -8.51,P=0.000) in CNE1and CNE1 R cells. Conclusions The MDR1 expression was increased after a total dose of50 Gy irradiation,which may be related to the synchronous change of Bcl-2 and MMP7 genes.
ABSTRACT
Purpose To study NF-κB gene expression level in mouse hepatocellular carcinoma cell lines with differently lymphatic metastasis potentials and to discuss its roles in lymphatic metastasis.Methods Using real-time quantitative PCR, NF-κB gene expression level was detected in mouse hepatocellular carcinoma cell lines, including Hca-P with low lymphatic metastasis potential and Hca-F with high lymphatic metastasis potential.Results NF-κB mRNA expression in Hca-P and Hca-F cell lines were (1.41±0.48)×10~(-3),and (2.95±0.22)×10~(-3) (P<0.01),respectively.NF-κB mRNA expression levels were increased with metastasis potential.Conclusion NF-κB gene may play an important role in lymphatic metastasis of hepatocellular carcinoma.
ABSTRACT
BACKGROUND: Opiate drugs are widely used to control chronic cancer pain, which brings many adverse reactions. Transplantation of sodium alginate-polylysine-alginate microencapsulated bovine adrenal chromaffin cell (BCC) has reported to be used as chronic cancer pain controllers. However, the high price and poor strength of polylysine limited its clinical application. Chitosan is characterized by sufficient resource, low price and good biocompatibility, which is a substitute for polylysine.OBJECTIVE: To study the effect of xenotransplantation of sodium alginate-chitosan-alginate (ACA) microencapsulated BCC on patients with advanced cancer pain.DESIGN, TIME AND SETTING: A retrospective case analysis. All cases were obtained from Department of Oncology, Affiliated Zhongshan Hospital of Dalian University from January 2007 to December 2008.PARTICIPANTS: Totally 10 patients with advanced cancer, including 1 female and 9 males, aged 46-78 years. According to visual algetic mimic scale (VAS), 3 patients suffered moderate pain and 7 cases suffered severe pain.METHODS: Microencapsulation method was applied to encapsulate BCC with ACA membrane and transplant the microencapsulated BCC (5-7)×10~6 into the subarachnoids pace of 10 patients.MAIN OUTCOME MEASURES: The degree of pain release, duration of analgesic effect, as well as adverse reaction.RESULTS: All 10 patients had pain relief rapidly after transplantation in varying degrees. Complete pain relief was shown in 2 cases, medium relief in 1 case, slight relief in 4 cases. Slight irritation of cauda eguina was presented after transplantation, which could disappear within 3-5 days.CONCLUSION: Xenotransplantation of ACA microencapsulated BCC into the spinal subarachnoids pace of patients with cancer pain can produce analgesic effect promptly, significantly, and safely.