| Objective:Colorectal cancer is one of malignant cancer with high morbidity and mortality. As a major developing country, with aging, changeing of lifestyle and living environment, the morbidity and mortality rates of colorectal cancer showed a rising trend. Colorectal cancer is the disease resulting in deterioration of public health and increasing economic burden of families and nation. The main reason of deterioration and death in patients is unlimited proliferation of cancer cells and extensive self-invasion and metastasis. PINCH (Particularly interesting new cysteine-histidine rich protein, PINCH), found in 1994, is one of LIM domain protein family members, The protein functions as an adapter protein for signal transduction in the integrin and growth factor pathways by binding ILK (integrin-linked kinase, ILK) and Nck-2, The location of high expression is in the stromal of the edge of the cancer invasion and PINCH is considered as an independent prognostic indicator in colorectal cancer. MAC30(Meningioma-associated protein-30), found in1993, is one of transmembrane protein. The expression is different in various human cancers and distinguished with the normal tissue. The levels of expression in colorectal cancer is high, associated with prognosis, but the specific mechanism of MAC30 is not clear.In this research, RT-PCR(Reversed Transcript PCR) was used to determine the mRNA expression level of PINCH and MAC30 in the primary tumor, adjacent non-carcerous mucosa, normal mucosa of proximal and distal margin in order to demonstrate their effects on the development and prognosis of coloretcal cancer. The correlation of mRNA expression in primary tumor with clinicopathologic variables of the cancer was analysised. The correlation between PINCH and MAC30 mRNA expression in primary tumor is also analysis.Methods: Tissue blocks were obtained from 80 patients with primary colorectal adenocarcinoma who underwent surgical resection in the First Hospital of Hebei Medical University and Tangshan Workers Hospital in November 2008 ~ March 2010. The patients'clinicopathologic features, such as sex, age, tumor location, and Dukes'stage, were obtained from surgical and/or pathologic records at these Hospitals. All 80 patients underwent PINCH mRNA detection, 49 males, 31 females. The mean age was 66 years (range from 37 to 90 years). 55 patients underwent MAC30 mRNA detection, 35 males, 20 females. The mean age was 66 years (range from 38 to 90 years).The specimens were grouped by positions,including primary tumor, adjacent non-carcerous mucosa(adjacent 2cm), normal mucosa of proximal and distal margin.Results:1 The expression situation PINCH mRNA in primary tumor, adjacent non-carcerous mucosa(adjacent 2cm), normal mucosa of proximal and distal margin(Fig.1~2 Fig.4):The expression level of primary tumor, was obviously increased compared to adjacent non-carcerous mucosa(adjacent 2cm), normal mucosa of proximal and distal margin (P<0.05). The different among adjacent non-carcerous mucosa(adjacent 2cm), normal mucosa of proximal and distal margin is not significant difference in the statistics (P<0.05).2 The expression situation MAC30 mRNA in primary tumor, adjacent non-carcerous mucosa(adjacent 2cm), normal mucosa of proximal and distal margin(Fig.3 Fig.5):The expression level of primary tumor, was obviously increased compared to adjacent non-carcerous mucosa(adjacent 2cm), normal mucosa of proximal and distal margin (P<0.05). The different among adjacent non-carcerous mucosa(adjacent 2cm), normal mucosa of proximal and distal margin is not significant difference in the Statistics (P<0.05).3 The correlation between expression level of PINCH mRNA in primary tumor and clinicopathological features:(Table 1)PINCH mRNA expression are not related to gender. Male group correction(PINCH/β-actin) is 0.678±0.378, female group correction 0.772±0.459. There is no significant difference between them (P=0.319,>0.05).PINCH mRNA expression are not related to age. <60 group correction(PINCH/β-actin) is 0.720±0.429,≥60 group correction 0.699±0.404. There is no significant difference between them (P = 0.823,> 0.05).PINCH mRNA expression are not related to tumour location. Colon group correction(PINCH/β-actin) is 0.816±0.510, rectal group correction 0.635±0.293. There is no significant difference between them (P=0.74,> 0.05).PINCH mRNA expression are not related to histological type. Non-mucinous group correction(PINCH/β-actin) is 0.682±0.349, mucinous/signet-ring cell group correction 0.716±0.445. There is no significant difference between them (P = 0.319,> 0.05).PINCH mRNA expression are not related to lymph node status. Non-metastasis group correction(PINCH/β-actin) is 0.689±0.407, metastasis group correction 0.742±0.419. There is no significant difference between them (P =0.569,>0.05).PINCH mRNA expression are not related to invasive depth. Intra-bowel wall group correction(PINCH/β-actin) is 0.544±0.324,ultra-bowel wall group correction 0.747±0.417. There is no significant difference between them (P =0.191,> 0.05).PINCH mRNA expression are not related to grade of differentiation. Better group correction(PINCH/β-actin) is 0.661±0.369, worse correction 0.794±0.439.There is no significant difference between them (P =0.191, >0.05).PINCH mRNA expression are related to Dukes'stage. Dukes'A+B+C stage group correction(PINCH/β-actin) is 0.736±0.424, Dukes'D stage group correction 0.525±0.200. There is significant difference between them ( P = 0.028,< 0.05).PINCH expression was not related to patient's gender, age, tumour location, histological type, differentiation, invasion depth, lymph node status , but related to Dukes'stage.4 The correlation between expression level of MAC30 mRNA in primary tumor and clinicopathological features (Table 1):MAC30 mRNA expression are not related to gender. male group correction(MAC30/β-actin) is 0.532±0.358, female group correction 0.544±0.301.There is no significant difference between them (P=0.895,>0.05).MAC30 mRNA expression are not related to age. <60 group correction(MAC30/β-actin) is 0.568±0.298,≥60 group correction 0.520±0.356. There is no significant difference between them (P =0.823,>0.05).MAC30 mRNA expression are not related to Dukes'stage. Dukes'A+B+C stage group correction(MAC30/β-actin) is 0.547±0.340, Dukes'D stage group correction 0.451±0.313. There is no significant difference between them ( P = 0.516, > 0.05).MAC30 mRNA expression are not related to histological type. Non-mucinous group correction(MAC30/β-actin) is 0.504±0.321, mucinous/signet-ring cell group correction 0.620±0.378. There is no significant difference between them (P =0.253,> 0.05).MAC30 mRNA expression are not related to lymph node status tumour location. Non-metastasis group correction (MAC30/β-actin) is 0.541±0.312, metastasis group correction 0.529±0.471. There is no significant difference between them (P = 0.74, >0.05).MAC30 mRNA expression are not related to grade of differentiation. Better group correction(MAC30/β-actin) is 0.502±0.311, worse correction 0.617±0.313. There is no significant difference between them (P = 0.371, > 0.05).MAC30 mRNA expression are related to tumour location and invasive depth. Colon group correction(MAC30/β-actin) is 0.677±0.419, rectal group correction group correction 0.412±0.162. There is significant difference between them (P=0.005,<0.05). Intra-bowel wall group correction(MAC30/β-actin) is 0.404±0.115, ultra-bowel wall group correction group correction 0.571±0.364. There is significant difference between them (P = 0.014,< 0.05).MAC30 expression was not related to patient's gender, age, Dukes'stage histological type, differentiation, lymph node status , but related to tumour location and invasion depth.5 Correlation analysis between PINCH and MAC30 mRNA in colorectal cancer:The expression between PINCH and MAC30 mRNA had a positive correlation with r = 0.275, P = 0.048 (Fig.6).The sample is divided into PINCH mRNA high expression and low expression group by arithmetic average of PINCH mRNA expression.The relationship of MAC30 mRNA expression between the two groups is analysed by the Mann-Whitney Test. MAC30 mRNA expression in PINCH mRNA high expression group was 0.673±0.386, mean rank 32.35.MAC30 mRNA expression in PINCH mRNA low expression group was 0.484±0.307, mean rank 23.66.The result of non-parametric statistics,Z=-1.941 , P=0.052 (P>0.05),suggested there was an increasing tendency in MAC30 mRNA expression relating to PINCH mRNA expression(Fig.7).Samely the sample is divided into MAC30 mRNA high expression and low expression group by arithmetic average of MAC30 mRNA expression.The relationship of PINCH mRNA expression between the two groups is analysed by the Mann-Whitney Test.PINCH mRNA expression in MAC30 mRNA high expression group was 0.823±0.502, mean rank 30.88.PINCH mRNA expression in MAC30 mRNA low expression group was 0.635±0.429, mean rank 21.2.The result of non-parametric statistics, Z=-2.086,P=0.037 (P<0.05), suggested there was significant difference between the two groups,and there was an increasing tendency in PINCH mRNA expression relating to MAC30 mRNA expression(Fig.8). Conclusions:PINCH and MAC30 are related with the development and invasiveness of colorectal cancer. The overexpression of PINCH and MAC30 may be an important indicater suggesting early infiltration. |
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