Association of inflammatory markers with the disease & mutation status in pancreatic cancer

Background & objectives: Inflammation has been studied to be an important contributory factor to carcinogenesis through pro-inflammatory markers such as interleukin (IL)-6 and C-reactive protein (CRP). Furthermore, K-ras mutation is an important genetic alteration in the pathogenesis of pancreatic cancer. This study aimed to compare these inflammatory markers in pancreatic ductal adenocarcinoma (PDAC) with the diseased and healthy controls (HCs) and to check for any association between IL-6 and CRP serum levels with the disease status, survival and K-ras mutation status of PDAC patients. Methods: The study included 135 PDAC, 25 chronic pancreatitis (CP) patients and 25 HCs. The serum levels of IL-6 and CRP were detected by enzyme-linked immunosorbent assay and K-ras mutations were detected by polymerase chain reaction-restriction fragment length polymorphism technique. Results: The serum levels of both these markers were elevated in PDAC cases than that in HCs. High IL-6 levels and higher CRP levels were found to be associated with locally advanced disease, lymphatic invasion, metastasis and advanced stage of the PDAC. In patients with unresectable PDAC, higher IL-6 levels were found to be associated with the presence of K-ras mutations. Interpretation & conclusions: Higher IL-6 and CRP levels in patients with advanced PDAC suggest an important role of these inflammatory markers in tumour progression. Furthermore, the association of mutations in the K-ras gene with serum IL-6 indicates cross-talks that may contribute to the progression of the PDAC.

Precision Therapy of Metastatic Colorectal Cancer with RAS Mutation / 肿瘤防治研究

Metastatic colorectal cancer (mCRC) is a clinical and molecular heterogeneous disease. Currently, for mCRC, extended rat sarcoma (RAS) testing is recommended in routine clinical practice before any treatment. RAS mutational status is significantly associated with the outcome of patients and strongly predictive for anti-EGFR-targeted therapy. However, specific treatments for RAS target are not yet available. Previous studies have shown that direct inhibition of RAS proteins has limited clinical benefits. Recently, a promising drug, AMG-510, which can directly inhibit KRAS G12C has been reported; however, it needs further confirmation. In the past few years, important advances have also been made in approaches designed to indirectly target RAS by inhibiting RAS effectors, multi-target combination strategies and immunotherapy. They are expected to be effective treatments for RAS target. This article summarizes the precision treatment of RAS-mutant mCRC.

Inhibitory effects of CIK on lung cancer A549 cells intensified by chemotherapyinduced damaged associated molecular pattern / 中国肿瘤生物治疗杂志

@# Objective: To investigate the effect of DAMP (damaged associated molecular pattern) on the inhibition of RAS-mutant A549 lung adenocarcinoma cells by CIK cells and its mechanism. Methods: Human peripheral blood mononuclear cells were isolated in vitro and CIK cells were cultured. A549 cells were treated with cisplatin (DDP) and doxorubicin (ADM) alone or in combination, and the morphology of A549 cells was observed under a microscope. The supernatant of A549 cells was co-cultured with CIK cells. Flow cytometry was used to detect the CIK cell immunophenotype after co-culture. MTT assay was used to detect the inhibition of A549 lung cancer cell proliferation induced by A549 cell supernatant. The concentration of chemotherapeutic drugs kills A549 cell supernatant CRT, ATP, HMGB1 content. Results: Low-level chemotherapeutic drugs showed more immunogenic death characteristics after killingA549 cells. The ratio of CD8+ and CD56+ in CIK cells was significantly higher than that in control CIK cells (P<0.05). The inhibition rate of CIK cells induced byA549 cells after injury onA549 lung adenocarcinoma cells was significantly higher than that of the same dose chemotherapy group [DDP group (31.34±1.51)% vs (5.97±1.74)%, ADM group (45.46±1.78)% vs (6.22±1.34)%, DDP+ ADM group (45.78±1.14)% vs (11.94±3.11)%, all P<0.05], and low-mass chemotherapeutic agents killed C549 induced by A549 cell supernatant on A549 The inhibition rate of the cells was higherthan that of the supernatant induced by the higher concentration of chemotherapeutic drugs (all P<0.05). The level of CRT,ATP, and HMGB1 in immunogenicity-related molecules in the supernatant ofA549 cells was significantly increased by low-concentration chemotherapy drugs (all P<0.05). In the low-concentration group, the supernatant-induced inhibition of the proliferation of A549 lung adenocarcinoma cells increased with the increase of CRT, ATP, and HMGB1 levels. Conclusion: The combination of lower concentration of DDP and ADM alone or in combination could more easily induce the immunogenic death of A549 cells and release higher levels of DAMP molecules, which could promote the inhibitory effect of CIK on lung cancerA549 cells.

Mutation of k-ras mutation and evaluation of anti-EGFR monoclonal antibody combined with chemotherapy in Chinese colorectal cancer patients / 肿瘤研究与临床

Objective To observe the k-ras mutation rate of colorectal cancer in China, and assess the effect and toxicity in advanced colorectal cancer (CRC) patients (pts) receiving chemotherapy combined with monoclonal antibody against Epidermal Growth Factor Receptor (EGFR). Methods The k-ras mutation of 139 samples collected from our hospital were tested by pyrophosphoric acid sequencing. Twenty-three advanced colorectal cancer patients were treated with chemotherapy combined with anti-EGFR monoclonal antibody, including 3 initial treated and 20 retreated. In the total 23 patients, 18 were treated with cetuximab and chemotherapy, and S were treated with nimotuzumab and chemotherapy. Cetuximab was taken with 400 mg/m2 first time, and then 250 mg/m2 every week. Nimotuzumab was taken with 400 mg first time, and then 200 mg every week. Eight patients of the total 23 received anti-EGFR monoclonal antibody combined with irinotecan, 12 with FOLFIRI, 3 with FOLFOX4. Results k-ras mutation type (mt) was detected in 39.6 % (55/139) of pts, k-ras wild type (wt) was 60.4 % (94/139). In 22 effect evaluable patients, 5 received PR and 9 SD, and RR and DCR were 22.7 % and 63.6 %, respectively. TTP was 124 days. Thirteen of the 22 patients tested k-ras mutation, of the 11 k-ras mt pts, 4 received PR, 4 SD and 3 PD. Two patients of k-ras mt received PD after 2 cycles treatment. Acneform eruptions were observed in 15 patients of 18 pts who received cetuximab and paronychia in 3 pts. Eruption or paronychia was not observed in all patients who received nimotuzumab. Grade 3 hypersensitivity were occured in 2 patients with cetuximab, and one of them alternated nimotuzumab in next cycle didn't get hypersensitivity any more. Conclusion The mutation rate of k-ras in Chinese colorectal cancer patients was similar with westerners. The effect of cetuximab combined with chemotherapy on advanced colorectal cancer was reliable. Nimotuzumab combined with chemotherapy worth to be studied further because of the promising effect and mild toxicity.

The Clinical Significance of Transforming Growth Factor (TGF) beta1, TGF beta Receptor beta II, p53 Protein and K-ras Point Mutation in Pancreatic Cancer

PURPOSE: Many cancers, including pancreatic cancer, harbor defects in TGF beta signaling and are resistant to TGF beta mediated growth inhibition. In addition, the expression of the p53 gene and mutations in K-ras might play an important role in the multistep carcinogenesis of pancreatic cancer. This study examined the expression level of TGF beta 1, TGF beta receptorII (T beta RII), p53 protein and K-ras mutation in pancreatic cancer, along with their role and clinical significance. METHODS: The overexpression of TGF beta 1, T beta RII and p53 protein was evaluated using an immunohistochemical assay. The K-ras mutation was analyzed by PCR-RFLP in the surgical resected pancreatic tissue from 26 pancreatic ductal adenocarcinomas and 5 normal pancreases. RESULTS: Immunohistochemical analysis of TGF beta 1 and T beta RII revealed positive immunostaining in 73.1% and 76.9% of the tumors, respectively, which were significantly higher than the normal pancreas (P=0.008). The p53 protein was positive in none of the 5 normal ducts and 16 out of 26 (61.5%) pancreatic carcinoma specimens. The K-ras mutation was positive in none of the 5 normal ducts, and in 20 of the 26 pancreatic carcinoma specimens (76.9%). The presence of TGF beta1 and T beta RII in the cancer samples was significantly associated with node metastasis, advanced tumor stage (P<0.01), and a short survival time (P<0.05). The p53-positive pancreatic cancers showed a significantly lower survival rate than those with p53-negative tumors (P<0.05). There was no correlation between K-ras mutations and the survival rates. CONCLUSION: The detection of K-ras mutations and TGF beta 1, T beta RII and p53 protein overexpression can predict the prognosis of pancreatic carcinoma patients.

Detection of K-ras mutation in DNA extracted from the plasma of patients with colorectal cancer / 中华消化杂志

Objective The purpose of this study was to determine the value of K-ras mutation in DNA extracted from the plasma as clinical indicator of colorectal cancer. Methods Point mutation at codon 12 of K-ras gene was assayed by polymerase chain reaction of sequence-specific primers in DNA extracted from the plasma and tumors from 32 patients with colorectal cancer. The mutation was further confirmed by dideoxy-mediated chain-termination method of DNA sequencing. Results Fourteen patients (44%) had a codon 12 K-ras mutation within their primary tumors and identical mutations were found in the plasma DNA of 13 patients (93%). Mutant DNA was not detected in the plasma specimens of 18 patients whose tumors tested negative for K-ras alterations or in 5 healthy control subjects. Conclusion Preliminary results suggest the detection of K-ras mutations in circulating DNA extracted from the plasma specimens may have some clinical uti- lity in the detection of colorectal cancer.

The Relationship of HPV Infection with Ki-ras and p53 Mutation in Uterine Cervical Carcinomas / 대한산부인과학회잡지

OBJECTIVE: Human papillomavirus(HPV) is an important pathogenetic agent in the uterine cervical carcinoma. It has been known that E6 and E7 proteins of high risk HPV viruses abolish the action of tumor suppressor genes, Rb and p53, resulting in cellular proliferation. This study was designed to determine the incidence of Ki-ras and p53 mutations in uterine cervical carcinomas and evaluate the results of mutations according to the status of HPV infection. METHODS: Paraffin-embedded tissue samples obtained from 42 CIN III and 70 invasive carcinomas of the uterine cervix(56 squamous, 7 adenosquamous and 7 adenocarcinomas). Ki-ras codon 12 mutation was analysed by PCR-RFLP method and p53 exons 5, 6, 7 and 8 mutations by PCR-SSCP. The presence of HPV was detected by PCR using consensus primers for high risk viruses(HPV 16, 18, 31, 33, 35, 52b, and 58) and low risk viruses(HPV 6 and 11). RESULTS: HPV viruses were detected in 83.3% of CIN III and 90% of invasive carcinomas; 71.4% of adenocarcinomas, 91.7% of squamous cell carcinomas and 100% of adenosquamous carcinomas. Ki-ras codon 12 mutation was detected in 4.7% of CIN III and 11.4% of invasive carcinomas, and p53 mutations in 4.8% of CIN III and 7.1% of invasive carcinomas. There was no significant difference of Ki-ras mutation between HPV-positive group(9.2%) and HPV-negative group(7.1%). However, p53 mutation was more frequent in HPV-negative group(28.6%) than HPV-positive group(3.1%). (p=0.0002) CONCLUSIONS: HPV is an agent which is detected in high frequency of uterine cervical carcinomas and p53 mutation may have a role in the HPV-negative uterine cervical carcinomas. However, the relationship between Ki-ras gene and HPV in uterine cervical carcinogenesis remains to be defined.

Effects of the Mutation of the p53 Tumor Suppressor Gene and the K-ras Gene on Clinical Manifestation in Non-small Cell Lung Cancer / 대한내과학회지

OBJECTIVE: A multistep process of gene alterations is required for tumor formation. p53 gene mutation is the most frequent and K-ras gene mutation places second in the gene abnormalities of non-small cell lung cancer (NSCLC). The effect by the mutations of the p53 and ras genes on clinical manifestation is still highly controversial. Little is known about the interaction between them in NSCLC. The present study was designed to investigate the effect by the mutations of the p53 tumor suppressor gene and K-ras oncogene on clinical manifestation, and the interaction between the mutations of two genes in the Korean NSCLC. METHODS: Fifty-eight patients were enrolled in this study who had been diagnosed as having NSCLC from stageI to stage III. They all had been alive for more than one month without any complication after curative resection. The paraffin-embedded lung tissues after resection were used to investigate the p53 expression by immunohistochemical staining, the mutations of the p53 and K-ras genes by polymerase chain reactionsingle strand conformation polymorphism (PCR-SSCP) and nucleotide sequencing. RESULTS: p53 protein was overexpressed in 25.9% by immunohistochemical staining. Overexpression was significantly more frequent in epidermoid carcinoma (p=0.01634). But there was no significant difference between the overexpression group and the negative expression group according to stage and survival. By PCR-SSCP analysis, the mobility shift of the p53 gene was found in 29.1%. There was no significant difference between the groups with and without mobility shift according to cell type, stage and survival. By nucleotide sequencing, p53 gene mutation was 37.9%. The locations of mutation were dispersed among numerous codons and the modes of mutation were also diverse. There was also no significant difference between the groups with and without mutation according to cell type, stage and survival. K-ras gene mutation was 24.1% and only in codon 12 by nucleotide sequencing. Although there was no significant difference between the groups with and without mutation according to cell type or stage, K-ras gene mutation carried a significantly worse prognosis in NSCLC (overall survival p=0.0391, disease-free survival p=0.0318). When the patients were divided into 4 groups according to p53 gene mutation and K-ras gene mutation, there was also no significant difference among any group according to cell type or stage. The prognosis became worse if K-ras gene mutation accompanied (overall survival p=0.0021, disease- free survival p=0.0166). Only the stage (p=0.0313) and K-ras gene mutation (p=0.0457) were significant prognostic factors by Cox regression test. An analysis in stage III showed the significantly shorter survival period in the patients with K-ras gene mutation. K-ras gene mutation, therefore, was confirmed as the independently significant prognostic factor separately from stage. CONCLUSION: p53 gene mutation had no clinical or prognostic significance because of scattered locations and diverse modes of mutation in contrast to K-ras gene mutation, which had a significantly negative effect on the prognosis of NSCLC. p53 and K-ras gene mutations were apparently independent genetic alterations which played different roles in the clinical manifestation and prognosis of NSCLC.

Correlation between K-ras gene mutation and non-small cell lung cancer / 西安交通大学学报(医学版)

Objective To explore the clinical significance of K-ras mutation in non-small cell lung cancer. Methods A K-ras codon 12 point mutation was detected by restriction fragment length polymorphisms combined with polymerase chain reaction in 41 tissues from non-small cell lung cancer (NSCLC), 21 normal tissues from nearby tumor and 21 lymph nodes from nearby bronchi. Results K-ras mutation was detected in 10 of 41 NSCLC((24.39%)). There was no relationship between K-ras mutation and the age, sex, differentiation of tumor, tumor volume,histologic type and tumor stage. K-ras mutation was more common in smoking group than in non-smoking group, but the statistical comparison was not significant. There was a strong link between cigarette smoking and K-ras point mutation in lung adenocarcinoma, but there was no relation between cigarette smoking amount and K-ras mutation. K-ras mutation was found in 4 of 21 metastatic lymph notes. The mortality in positive K-ras mutation was significantly higher than that in negative K-ras mutation(P